Cyclobutyl purine derivative or salt thereof

ABSTRACT

(in the formula, R1 represents a halogen atom, an amino group which may be substituted, a C1-6 alkoxy group which may be substituted, a hydroxyl group which may be protected, or the like; R2 represents a hydrogen atom or an amino protecting group; R3 represents a C1-20 alkoxy group which may be substituted, an aryloxy group which may be substituted, an amino group which may be substituted, or the like; R4 represents a C1-20 alkoxy group which may be substituted, an aryloxy group which may be substituted, an amino group which may be substituted, or the like); or a salt thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of PCT International Application No.PCT/JP2019/029448 filed on Jul. 26, 2019, which claims priority under 35U.S.C § 119(a) to Japanese Patent Application No. 2018-141749 filed onJul. 27, 2018. Each of the above application(s) is hereby expresslyincorporated by reference, in its entirety, into the presentapplication.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention relates to a cyclobutyl purine derivative or asalt thereof and an anti-adenoviral agent containing the same.

2. Description of the Related Art

Adenovirus is a double-stranded linear DNA virus and is a causativevirus that causes various pathological conditions such as respiratoryinfection, pharyngoconjunctival fever, epidemic keratoconjunctivitis,hepatitis, gastroenteritis, cystitis, and encephalitis.

Although there is no pharmaceutical product approved as ananti-adenoviral agent, for example, cidofovir is known to be clinicallyeffective (Antiviral Research, vol. 71, pp. 172 to 180, 2006). On theother hand, cidofovir is known to be toxic upon systemic administrationand ocular instillation administration (Antimicrobial Agents andChemotherapy, vol. 59(7), pp. 3718 to 3725, 2015, and Drugs, vol. 71(3),pp. 331 to 347, 2011).

On the other hand, cyclobutyl purine derivatives are expected to beapplied to various pharmaceutical products, and for example, compoundshaving an anti-herpesvirus effect and an anti-human immunodeficiencyvirus activity are known (JP1990-131473A (JP-H02-131473A)).

SUMMARY OF THE INVENTION

An object of the present invention is to provide a compound exhibitingan excellent drug efficacy against adenovirus and an excellentanti-adenoviral agent.

As a result of extensive studies, the present inventors have found thata compound represented by General Formula [1] or a salt thereof exhibitsan excellent drug efficacy against adenovirus. The present invention hasbeen completed based on these findings.

The present invention provides the following.

(1) A compound represented by General Formula [1]:

in the formula,

R¹ represents a halogen atom, an amino group which may be substituted, aC₁₋₆ alkoxy group which may be substituted, a C₃₋₈ cycloalkoxy groupwhich may be substituted, a C₁₋₆ alkylthio group which may besubstituted, a hydroxyl group which may be protected, or a thiol groupwhich may be protected;

R² represents a hydrogen atom or an amino protecting group;

R³ represents a C₁₋₂₀ alkoxy group which may be substituted, a C₃₋₈cycloalkoxy group which may be substituted, a C₁₋₂₀ alkylthio groupwhich may be substituted, an aryloxy group which may be substituted, aheterocyclic ring group which may be substituted, a heterocyclic oxygroup which may be substituted, an amino group which may be substituted,or —O—P(O)(OH)—O—PO₃H; and

R⁴ represents a C₁₋₂₀ alkoxy group which may be substituted, a C₃₋₈cycloalkoxy group which may be substituted, a C₁₋₂₀ alkylthio groupwhich may be substituted, an aryloxy group which may be substituted, aheterocyclic ring group which may be substituted, a heterocyclic oxygroup which may be substituted, an amino group which may be substituted,or a hydroxyl group which may be protected; or

R³ and R⁴, together with a phosphorus atom to which R³ and R⁴ arebonded, may be combined to form a 5- to 10-membered nitrogen- andphosphorus-containing heterocyclic ring which may be substituted, a 5-to 10-membered oxygen- and phosphorus-containing heterocyclic ring whichmay be substituted, or a 5- to 10-membered nitrogen-, oxygen-, andphosphorus-containing heterocyclic ring which may be substituted,provided that R² represents a hydrogen atom in a case where R³represents —O—P(O)(OH)—O—PO₃H; or

a salt thereof.

(2) The compound according to (1), in which R² is a hydrogen atom; orthe salt thereof.

(3) The compound according to (1) or (2), in which R¹ is a halogen atom,an amino group which may be substituted with one or more substituentsselected from Substituent group A, a C₁₋₆ alkoxy group which may besubstituted with one or more substituents selected from Substituentgroup A, a C₃₋₈ cycloalkoxy group which may be substituted with one ormore substituents selected from Substituent group A, a C₁₋₆ alkylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group A, a hydroxyl group which may be protected, or athiol group which may be protected; or the salt thereof.

Substituent Group A:

a halogen atom; a hydroxyl group which may be protected; a cyano group;a nitro group; a carbamoyl group; an oxo group; a C₁₋₆ alkyl group whichmay be substituted with one or more substituents selected fromSubstituent group B; a C₁₋₆ alkyldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₂₋₆ alkenyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₂₋₆ alkynyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a C₃₋₈ cycloalkyl group which may be substitutedwith one or more substituents selected from Substituent group B; a C₃₋₈cycloalkyldisulfanyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₁₋₆ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a C₁₋₆ alkoxycarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₃₋₈ cycloalkoxycarbonyl group which may be substituted withone or more substituents selected from Substituent group B; an arylgroup which may be substituted with one or more substituents selectedfrom Substituent group B; an aryldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a heterocyclic ring group which may be substituted with one ormore substituents selected from Substituent group B; a heterocyclic oxygroup which may be substituted with one or more substituents selectedfrom Substituent group B; an acyloxy group which may be substituted withone or more substituents selected from Substituent group B; an acylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group B; an aminocarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B; and an aminocarbonylthio group which may be substituted withone or more substituents selected from Substituent group B.

Substituent Group B:

a halogen atom; a hydroxyl group; a cyano group; a nitro group; acarboxyl group; a carbamoyl group; a hydroxymethyl group; a C₁₋₆ alkylgroup; a C₂₋₆ alkenyl group; a C₂₋₆ alkynyl group; a C₃₋₈ cycloalkylgroup; a C₁₋₆ alkoxy group; a C₁₋₆ alkoxycarbonyl group; a C₃₋₈cycloalkoxycarbonyl group; an ar-C₁₋₆ alkoxycarbonyl group; an arylgroup; an aryloxy group; a heterocyclic oxy group which may besubstituted with one or more substituents selected from a hydroxyl groupand a hydroxymethyl group; an ar-C₁₋₆ alkoxy group; and an acyloxygroup.

(4) The compound according to (3), in which R¹ is a halogen atom, anamino group which may be substituted with one or more substituentsselected from Substituent group A, a C₁₋₆ alkoxy group which may besubstituted with one or more substituents selected from Substituentgroup A, a hydroxyl group which may be protected, or a thiol group whichmay be protected; or the salt thereof.

(5) The compound according to (3), in which R¹ is a halogen atom, anamino group which may be substituted with one or more substituentsselected from Substituent group A, a C₁₋₆ alkoxy group which may besubstituted with one or more substituents selected from Substituentgroup A, or a hydroxyl group; or the salt thereof.

(6) The compound according to any one of (1) to (5), in which R³ is aC₁₋₂₀ alkoxy group which may be substituted with one or moresubstituents selected from Substituent group A, a C₃₋₈ cycloalkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, a C₁₋₂₀ alkylthio group which may be substitutedwith one or more substituents selected from Substituent group A, anaryloxy group which may be substituted with one or more substituentsselected from Substituent group A, a heterocyclic ring group which maybe substituted with one or more substituents selected from Substituentgroup A, a heterocyclic oxy group which may be substituted with one ormore substituents selected from Substituent group A, an amino groupwhich may be substituted with one or more substituents selected fromSubstituent group A, or —O—P(O)(OH)—O—PO₃H; or the salt thereof.

Substituent Group A:

a halogen atom; a hydroxyl group which may be protected; a cyano group;a nitro group; a carbamoyl group; an oxo group; a C₁₋₆ alkyl group whichmay be substituted with one or more substituents selected fromSubstituent group B; a C₁₋₆ alkyldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₂₋₆ alkenyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₂₋₆ alkynyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a C₃₋₈ cycloalkyl group which may be substitutedwith one or more substituents selected from Substituent group B; a C₃₋₈cycloalkyldisulfanyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₁₋₆ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a C₁₋₆ alkoxycarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₃₋₈ cycloalkoxycarbonyl group which may be substituted withone or more substituents selected from Substituent group B; an arylgroup which may be substituted with one or more substituents selectedfrom Substituent group B; an aryldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a heterocyclic ring group which may be substituted with one ormore substituents selected from Substituent group B; a heterocyclic oxygroup which may be substituted with one or more substituents selectedfrom Substituent group B; an acyloxy group which may be substituted withone or more substituents selected from Substituent group B; an acylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group B; an aminocarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B; and an aminocarbonylthio group which may be substituted withone or more substituents selected from Substituent group B.

Substituent Group B:

a halogen atom; a hydroxyl group; a cyano group; a nitro group; acarboxyl group; a carbamoyl group; a hydroxymethyl group; a C₁₋₆ alkylgroup; a C₂₋₆ alkenyl group; a C₂₋₆ alkynyl group; a C₃₋₈ cycloalkylgroup; a C₁₋₆ alkoxy group; a C₁₋₆ alkoxycarbonyl group; a C₃₋₈cycloalkoxycarbonyl group; an ar-C₁₋₆ alkoxycarbonyl group; an arylgroup; an aryloxy group; a heterocyclic oxy group which may besubstituted with one or more substituents selected from a hydroxyl groupand a hydroxymethyl group; an ar-C₁₋₆ alkoxy group; and an acyloxygroup.

(7) The compound according to (6), in which R³ is a C₁₋₂₀ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, a C₁₋₂₀ alkylthio group which may be substitutedwith one or more substituents selected from Substituent group A, anaryloxy group which may be substituted with one or more substituentsselected from Substituent group A, an amino group which may besubstituted with one or more substituents selected from Substituentgroup A, or —O—P(O)(OH)—O—PO₃H; or the salt thereof.

(8) The compound according to (6), in which R³ is a C₁₋₂₀ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, a C₁₋₂₀ alkylthio group which may be substitutedwith one or more substituents selected from Substituent group A, anaryloxy group which may be substituted with one or more substituentsselected from Substituent group A, or —O—P(O)(OH)—O—PO₃H; or the saltthereof.

(9) The compound according to any one of (1) to (8), in which R⁴ is aC₁₋₂₀ alkoxy group which may be substituted with one or moresubstituents selected from Substituent group A, a C₃₋₈ cycloalkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, a C₁₋₂₀ alkylthio group which may be substitutedwith one or more substituents selected from Substituent group A, anaryloxy group which may be substituted with one or more substituentsselected from Substituent group A, a heterocyclic ring group which maybe substituted with one or more substituents selected from Substituentgroup A, a heterocyclic oxy group which may be substituted with one ormore substituents selected from Substituent group A, an amino groupwhich may be substituted with one or more substituents selected fromSubstituent group A, or a hydroxyl group which may be protected; or thesalt thereof.

Substituent Group A:

a halogen atom; a hydroxyl group which may be protected; a cyano group;a nitro group; a carbamoyl group; an oxo group; a C₁₋₆ alkyl group whichmay be substituted with one or more substituents selected fromSubstituent group B; a C₁₋₆ alkyldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₂₋₆ alkenyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₂₋₆ alkynyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a C₃₋₈ cycloalkyl group which may be substitutedwith one or more substituents selected from Substituent group B; a C₃₋₈cycloalkyldisulfanyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₁₋₆ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a C₁₋₆ alkoxycarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₃₋₈ cycloalkoxycarbonyl group which may be substituted withone or more substituents selected from Substituent group B; an arylgroup which may be substituted with one or more substituents selectedfrom Substituent group B; an aryldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a heterocyclic ring group which may be substituted with one ormore substituents selected from Substituent group B; a heterocyclic oxygroup which may be substituted with one or more substituents selectedfrom Substituent group B; an acyloxy group which may be substituted withone or more substituents selected from Substituent group B; an acylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group B; an aminocarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B; and an aminocarbonylthio group which may be substituted withone or more substituents selected from Substituent group B.

Substituent Group B:

a halogen atom; a hydroxyl group; a cyano group; a nitro group; acarboxyl group; a carbamoyl group; a hydroxymethyl group; a C₁₋₆ alkylgroup; a C₂₋₆ alkenyl group; a C₂₋₆ alkynyl group; a C₃₋₈ cycloalkylgroup; a C₁₋₆ alkoxy group; a C₁₋₆ alkoxycarbonyl group; a C₃₋₈cycloalkoxycarbonyl group; an ar-C₁₋₆ alkoxycarbonyl group; an arylgroup; an aryloxy group; a heterocyclic oxy group which may besubstituted with one or more substituents selected from a hydroxyl groupand a hydroxymethyl group; an ar-C₁₋₆ alkoxy group; and an acyloxygroup.

(10) The compound according to (9), in which R⁴ is a C₁₋₂₀ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, a C₁₋₂₀ alkylthio group which may be substitutedwith one or more substituents selected from Substituent group A, anaryloxy group which may be substituted with one or more substituentsselected from Substituent group A, an amino group which may besubstituted with one or more substituents selected from Substituentgroup A, or a hydroxyl group which may be protected; or the saltthereof.

(11) The compound according to (9), in which R⁴ is a C₁₋₂₀ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, an amino group which may be substituted with one ormore substituents selected from Substituent group A, or a hydroxyl groupwhich may be protected; or the salt thereof.

(12) The compound according to any one of (1) to (5), in which a ringformed by combining R³ and R⁴ together with a phosphorus atom to whichR³ and R⁴ are bonded is a 5-to 10-membered oxygen- andphosphorus-containing heterocyclic ring which may be substituted; or thesalt thereof.

(13) A compound selected from methyl

(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(naphthalen-1-yloxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chlorophenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate,ethyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate,ethyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chlorophenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-bromophenoxy)phosphoyl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-chlorophenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-(pivaloylthio)ethoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-ethoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate,methyl2-((((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)amino)-2-methylpropanoate,(cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methylbis(pivaloyloxymethyl)phosphate, methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chloro-2-fluorophenoxy)phosphoryl)-L-alaninate,and methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(3-bromophenoxy)phosphoryl)-L-alaninate;or a salt thereof.

(14) An anti-adenoviral agent comprising:

-   -   the compound or salt thereof according to any one of (1) to        (13).

(A) A medicine for treating an adenovirus infection, comprising:

-   -   the compound or salt thereof according to any one of (1) to        (13).

(B) A method for suppressing adenovirus, comprising:

-   -   administering the compound or salt thereof according to any one        of (1) to (13) to a subject (preferably a mammal such as a        human)

(C) A method for treating an adenovirus infection, comprising:

-   -   administering the compound or salt thereof according to any one        of (1) to (13) to a subject (preferably a mammal such as a        human)

(D) The compound or salt thereof according to any one of (1) to (13),for use in suppressing adenovirus.

(E) The compound or salt thereof according to any one of (1) to (13),for use in the treatment of an adenovirus infection.

(F) Use of the compound or salt thereof according to any one of (1) to(13) for the production of an anti-adenoviral agent.

(G) Use of the compound or salt thereof according to any one of (1) to(13) for the production of a medicine for treating an adenovirusinfection.

The compound represented by General Formula [1] or a salt thereofaccording to an aspect of the present invention is useful as ananti-adenoviral agent. The compound represented by General Formula [1]or a salt thereof according to the aspect of the present invention isuseful as an agent for treating adenovirus.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the results of electrophoresis. An extension reaction ofDNA was evaluated by detecting a fluorescently labeled DNA.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Hereinafter, the present invention will be described in detail.

In the present invention, the individual terms have the followingmeanings, unless otherwise indicated.

The halogen atom refers to a fluorine atom, a chlorine atom, a bromineatom, or an iodine atom.

The C₁₋₆ alkyl group refers to a linear or branched C₁₋₆ alkyl groupsuch as a methyl group, an ethyl group, a propyl group, an isopropylgroup, a butyl group, a sec-butyl group, an isobutyl group, a tert-butylgroup, a pentyl group, an isopentyl group, a 2-methylbutyl group, a2-pentyl group, a 3-pentyl group, or a hexyl group.

The C₁₋₂₀ alkyl group refers to a linear or branched C₁₋₂₀ alkyl groupsuch as a methyl group, an ethyl group, a propyl group, an isopropylgroup, a butyl group, a sec-butyl group, an isobutyl group, a tert-butylgroup, a pentyl group, an isopentyl group, a 2-methylbutyl group, a2-pentyl group, a 3-pentyl group, a hexyl group, a 4-methylpentyl group,a 3-methylpentyl group, a 2-methylpentyl group, a 1-methylpentyl group,a 3,3-dimethylbutyl group, a 2,2-dimethylbutyl group, a1,1-dimethylbutyl group, a 1,2-dimethylbutyl group, a 1,3-dimethylbutylgroup, a 2,3-dimethylbutyl group, a 2-ethylbutyl group, a heptyl group,a 1-methylhexyl group, a 2-methylhexyl group, a 3-methylhexyl group, a4-methylhexyl group, a 5-methylhexyl group, a 1-propylbutyl group, a4,4-dimethylpentyl group, an octyl group, a 1-methylheptyl group, a2-methylheptyl group, a 3-methylheptyl group, a 4-methylheptyl group, a5-methylheptyl group, a 6-methylheptyl group, a 1-propylpentyl group, a2-ethylhexyl group, a 5,5-dimethylhexyl group, a nonyl group, a decylgroup, an undecyl group, a dodecyl group, a tridecyl group, a tetradecylgroup, a pentadecyl group, a hexadecyl group, a heptadecyl group, anoctadecyl group, a nonadecyl group, or an eicosanyl group.

The C₁₋₆ alkylthio group refers to a linear or branched C₁₋₆ alkylthiogroup such as a methylthio group, an ethylthio group, a propylthiogroup, an isopropylthio group, a butylthio group, an isobutylthio group,a sec-butylthio group, a tert-butylthio group, a pentylthio group, or ahexylthio group.

The C₁₋₂₀ alkylthio group refers to a linear or branched C₁₋₂₀ alkylthiogroup such as a methylthio group, an ethylthio group, a propylthiogroup, an isopropylthio group, a butylthio group, an isobutylthio group,a sec-butylthio group, a tert-butylthio group, a pentylthio group, ahexylthio group, a heptylthio group, an octylthio group, a nonylthiogroup, a decylthio group, an undecylthio group, a dodecylthio group, atridecylthio group, a tetradecylthio group, a pentadecylthio group, ahexadecylthio group, a heptadecylthio group, an octadecylthio, anonadecylthio group, or an eicosanylthio group.

The C₁₋₆ alkylsulfonyl group refers to a C₁₋₆ alkylsulfonyl group suchas a methylsulfonyl group, an ethylsulfonyl group, or a propylsulfonylgroup.

The C₁₋₆ alkylsulfonyloxy group refers to a C₁₋₆ alkylsulfonyloxy groupsuch as a methylsulfonyloxy group or an ethylsulfonyloxy group.

The C₁₋₆ alkyldisulfanyl group refers to a linear or branched C₁₋₆alkyldisulfanyl group such as a methyldisulfanyl group, anethyldisulfanyl group, a propyldisulfanyl group, an isopropyldisulfanylgroup, a butyldisulfanyl group, a sec-butyldisulfanyl group, anisobutyldisulfanyl group, a tert-butyldisulfanyl group, apentyldisulfanyl group, an isopentyldisulfanyl group, a2-methylbutyldisulfanyl group, a 2-pentyldisulfanyl group, a3-pentyldisulfanyl group, or a hexyldisulfanyl group.

The C₂₋₆ alkenyl group refers to a linear or branched C₂₋₆ alkenyl groupsuch as a vinyl group, an allyl group, a propenyl group, an isopropenylgroup, a butenyl group, an isobutenyl group, a 1,3-butadienyl group, apentenyl group, or a hexenyl group.

The C₂₋₆ alkynyl group refers to a linear or branched C₂₋₆ alkynyl groupsuch as an ethynyl group, a propynyl group, a butynyl group, a pentynylgroup, or a hexynyl group.

The C₃₋₈ cycloalkyl group refers to a C₃₋₈ cycloalkyl group such as acyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexylgroup, or a cycloheptyl group.

The C₃₋₈ cycloalkyldisulfanyl group refers to a C₃₋₈cycloalkyldisulfanyl group such as a cyclopropyldisulfanyl group, acyclobutyldisulfanyl group, cyclopentyldisulfanyl group, acyclohexyldisulfanyl group, or a cycloheptyldisulfanyl group.

The C₁₋₆ alkoxy group refers to a linear or branched C₁₋₆ alkyloxy groupsuch as a methoxy group, an ethoxy group, a propoxy group, an isopropoxygroup, a butoxy group, an isobutoxy group, a sec-butoxy group, atert-butoxy group, a pentyloxy group, or a hexyloxy group.

The C₁₋₂₀ alkoxy group refers to a linear or branched C₁₋₂₀ alkyloxygroup such as a methoxy group, an ethoxy group, a propoxy group, anisopropoxy group, a butoxy group, an isobutoxy group, a sec-butoxygroup, a tert-butoxy group, a pentyloxy group, a hexyloxy group, aheptyloxy group, an octyloxy group, a nonyloxy group, a decyloxy group,an undecyloxy group, a dodecyloxy group, a tridecyloxy group, atetradecyloxy group, a pentadecyloxy group, a hexadecyloxy group, aheptadecyloxy group, an octadecyloxy group, a nonadecyloxy group, or aneicosanyloxy group.

The C₁₋₆ alkoxy C₁₋₆ alkyl group refers to a C₁₋₆ alkyloxy C₁₋₆ alkylgroup such as a methoxymethyl group or a 1-ethoxyethyl group.

The C₁₋₆ alkoxycarbonyl group refers to a linear or branched C₁₋₆alkyloxycarbonyl group such as a methoxycarbonyl group, anethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonylgroup, a butoxycarbonyl group, an isobutoxycarbonyl group, asec-butoxycarbonyl group, a tert-butoxycarbonyl group, apentyloxycarbonyl group, or a hexyloxycarbonyl group.

The C₁₋₂₀ alkoxycarbonyl group refers to a linear or branched C₁₋₂₀alkyloxycarbonyl group such as a methoxycarbonyl group, anethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonylgroup, a butoxycarbonyl group, an isobutoxycarbonyl group, asec-butoxycarbonyl group, a tert-butoxycarbonyl group, apentyloxycarbonyl group, a hexyloxycarbonyl group, a heptyloxycarbonylgroup, an octyloxycarbonyl group, a nonyloxycarbonyl group, adecyloxycarbonyl group, an undecyloxycarbonyl group, adodecyloxycarbonyl group, a tridecyloxycarbonyl group, atetradecyloxycarbonyl group, a pentadecyloxycarbonyl group, ahexadecyloxycarbonyl group, a heptadecyloxycarbonyl group, anoctadecyloxycarbonyl group, a nonadecyloxycarbonyl group, or aneicosanyloxycarbonyl group.

The C₁₋₆ alkoxycarbonyloxy group refers to a linear or branched C₁₋₆alkyloxycarbonyloxy group such as a methoxycarbonyloxy group, anethoxycarbonyloxy group, a propoxycarbonyloxy group, anisopropoxycarbonyloxy group, a butoxycarbonyloxy group, anisobutoxycarbonyloxy group, a sec-butoxycarbonyloxy group, atert-butoxycarbonyloxy group, a pentyloxycarbonyloxy group, or ahexyloxycarbonyloxy group.

The C₃₋₈ cycloalkoxy group refers to a C₃₋₈ cycloalkyloxy group such asa cyclopropoxy group, a cyclobutoxy group, a cyclopentyloxy group, acyclohexyloxy group, or a cycloheptyloxy group.

The C₃₋₈ cycloalkoxycarbonyl group refers to a C₃₋₈ cycloalkoxycarbonylgroup such as a cyclopropoxycarbonyl group, a cyclobutoxycarbonyl group,a cyclopentyloxycarbonyl group, a cyclohexyloxycarbonyl group, acycloheptyloxycarbonyl group, or a cyclooctyloxycarbonyl group.

The aryl group refers to a phenyl group or a naphthyl group.

The aryloxy group refers to a phenoxy group, a naphthalen-1-yloxy group,or a naphthalen-2-yloxy group.

The arylsulfonyl group refers to a benzenesulfonyl group, ap-toluenesulfonyl group, or a naphthalenesulfonyl group.

The arylsulfonyloxy group refers to a benzenesulfonyloxy group or ap-toluenesulfonyloxy group.

The aryldisulfanyl group refers to a phenyldisulfanyl group or anaphthyldisulfanyl group.

The ar-C₁₋₆ alkyl group refers to an ar-C₁₋₆ alkyl group such as abenzyl group, a diphenylmethyl group, a trityl group, a phenethyl group,a 2-phenylpropyl group, a 3-phenylpropyl group, or a naphthylmethylgroup.

The ar-C₁₋₆ alkoxy group refers to an ar-C₁₋₆ alkyloxy group such as abenzyloxy group, a diphenylmethoxy group, a trityloxy group, aphenethyloxy group, a 2-phenylpropoxy group, a 3-phenylpropoxy group, ora naphthylmethoxy group.

The ar-C₁₋₆ alkoxy C₁₋₆ alkyl group refers to an ar-C₁₋₆ alkyloxy C₁₋₆alkyl group such as a benzyloxymethyl group or a phenethyloxymethylgroup.

The ar-C₁₋₆ alkoxycarbonyl group refers to an ar-C₁₋₆ alkyloxycarbonylgroup such as a benzyloxycarbonyl group or a phenethyloxycarbonyl group.

The monocyclic nitrogen-containing heterocyclic ring group refers to amonocyclic nitrogen-containing heterocyclic ring group which containsonly a nitrogen atom as a heteroatom forming the ring, such as anaziridinyl group, an azetidinyl group, a pyrrolidinyl group, apyrrolinyl group, a pyrrolyl group, a piperidyl group, atetrahydropyridyl group, a dihydropyridyl group, a pyridyl group, ahomopiperidinyl group, an octahydroazocinyl group, an imidazolidinylgroup, an imidazolinyl group, an imidazolyl group, a pyrazolidinylgroup, a pyrazolinyl group, a pyrazolyl group, a piperazinyl group, apyrazinyl group, a pyridazinyl group, a pyrimidinyl group, ahomopiperazinyl group, a triazolyl group, or a tetrazolyl group.

The monocyclic oxygen-containing heterocyclic ring group refers to amonocyclic oxygen-containing heterocyclic ring group which contains onlyan oxygen atom as a heteroatom forming the ring, such as an oxetanylgroup, a tetrahydrofuranyl group, a furanyl group, a tetrahydropyranylgroup, a pyranyl group, a 1,3-dioxanyl group, or a 1,4-dioxanyl group.

The monocyclic sulfur-containing heterocyclic ring group refers to athienyl group.

The monocyclic nitrogen- and oxygen-containing heterocyclic ring grouprefers to a monocyclic nitrogen- and oxygen-containing heterocyclic ringgroup which contains only a nitrogen atom and an oxygen atom asheteroatoms forming the ring, such as an oxazolyl group, an isoxazolylgroup, an oxadiazolyl group, a morpholinyl group, or an oxazepanylgroup.

The monocyclic nitrogen- and sulfur-containing heterocyclic ring grouprefers to a monocyclic nitrogen- and sulfur-containing heterocyclic ringgroup which contains only a nitrogen atom and a sulfur atom asheteroatoms forming the ring, such as a thiazolyl group, an isothiazolylgroup, a thiadiazolyl group, a thiomorpholinyl group, a1-oxidothiomorpholinyl group, or a 1,1-dioxidothiomorpholinyl group.

The monocyclic heterocyclic ring group refers to a monocyclicnitrogen-containing heterocyclic ring group, a monocyclicoxygen-containing heterocyclic ring group, a monocyclicsulfur-containing heterocyclic ring group, a monocyclic nitrogen- andoxygen-containing heterocyclic ring group, or a monocyclic nitrogen- andsulfur-containing heterocyclic ring group.

The bicyclic nitrogen-containing heterocyclic ring group refers to abicyclic nitrogen-containing heterocyclic ring group which contains onlya nitrogen atom as a heteroatom forming the ring, such as an indolinylgroup, an indolyl group, an isoindolinyl group, an isoindolyl group, abenzimidazolyl group, an indazolyl group, a benzotriazolyl group, apyrazolopyridinyl group, a quinolyl group, a tetrahydroquinolinyl group,a quinolyl group, a tetrahydroisoquinolinyl group, an isoquinolinylgroup, a quinolizinyl group, a cinnolinyl group, a phthalazinyl group, aquinazolinyl group, a dihydroquinoxalinyl group, a quinoxalinyl group, anaphthyridinyl group, a purinyl group, a pteridinyl group, or aquinuclidinyl group.

The bicyclic oxygen-containing heterocyclic ring group refers to abicyclic oxygen-containing heterocyclic ring group which contains onlyan oxygen atom as a heteroatom forming the ring, such as a2,3-dihydrobenzofuranyl group, a benzofuranyl group, an isobenzofuranylgroup, a chromanyl group, a chromenyl group, an isochromanyl group, a1,3-benzodioxolyl group, a 1,3-benzodioxanyl group, or a1,4-benzodioxanyl group.

The bicyclic sulfur-containing heterocyclic ring group refers to abicyclic sulfur-containing heterocyclic ring group which contains only asulfur atom as a heteroatom forming the ring, such as a2,3-dihydrobenzothienyl group or a benzothienyl group.

The bicyclic nitrogen- and oxygen-containing heterocyclic ring grouprefers to a bicyclic nitrogen- and oxygen-containing heterocyclic ringgroup which contains only a nitrogen atom and an oxygen atom asheteroatoms forming the ring, such as a benzoxazolyl group, abenzisoxazolyl group, a benzoxadiazolyl group, a benzomorpholinyl group,a dihydropyranopyridyl group, a dioxolopyridyl group, a furopyridinylgroup, a dihydrodioxynopyridyl group, or a dihydropyridooxazinyl group.

The bicyclic nitrogen- and sulfur-containing heterocyclic ring grouprefers to a bicyclic nitrogen- and sulfur-containing heterocyclic ringgroup which contains a nitrogen atom and a sulfur atom as heteroatomsforming the ring, such as a benzothiazolyl group, a benzoisothiazolylgroup, or a benzothiadiazolyl group.

The bicyclic heterocyclic ring group refers to a bicyclicnitrogen-containing heterocyclic ring group, a bicyclicoxygen-containing heterocyclic ring group, a bicyclic sulfur-containingheterocyclic ring group, a bicyclic nitrogen- and oxygen-containingheterocyclic ring group, or a bicyclic nitrogen- and sulfur-containingheterocyclic ring group.

The spiro heterocyclic ring group refers to a spiro heterocyclic ringgroup which contains a nitrogen atom, an oxygen atom, or a sulfur atomas a heteroatom forming the ring, such as a 2-oxa-6-azaspiro[3.3]heptylgroup, a 1,4-dioxaspiro[4.5]decyl group, a 1-oxa-8-azaspiro[4.5]decylgroup, or a 1-thia-8-azaspiro[4.5]decyl group.

The bridged heterocyclic ring group refers to a bridged heterocyclicring group which contains a nitrogen atom, an oxygen atom, or a sulfuratom as a heteroatom forming the ring, such as a3-oxa-8-azabicyclo[3.2.1]octyl group, an 8-oxa-3-azabicyclo[3.2.1]octylgroup, or a quinuclidinyl group.

The heterocyclic ring group refers to a monocyclic heterocyclic ringgroup, a bicyclic heterocyclic ring group, a spiro heterocyclic ringgroup, or a bridged heterocyclic ring group.

The heterocyclic oxy group refers to a substituent in which an oxygenatom is bonded to a heterocyclic ring group such as pyrrolidinyloxy,piperidinyloxy, tetrahydrofuranyloxy, tetrahydropyranyloxy, ortetrahydrothiopyranyloxy.

The 5- to 10-membered nitrogen- and phosphorus-containing heterocyclicring refers to a 5- to 10-membered nitrogen- and phosphorus-containingheterocyclic ring which contains only a nitrogen atom and a phosphorusatom as heteroatoms forming the ring and which may be fused, such as1,3,2-diazaphospholidine, 1,3,2-diazaphosphinane,1,3,2-diazaphosphepane, or 1,3,2-diazaphosphocane.

The 5- to 10-membered oxygen- and phosphorus-containing heterocyclicring refers to a 5- to 10-membered oxygen- and phosphorus-containingheterocyclic ring which contains only an oxygen atom and a phosphorusatom as heteroatoms forming the ring and which may be fused, such as1,3,2-dioxaphospholane, 1,3,2-dioxaphosphinane, 1,3,2-dioxaphosphepane,1,3,2-dioxaphosphocane, benzo[d][1,3,2]dioxaphosphor, or4H-benzo[d][1,3,2]dioxaphosphinine.

The 5- to 10-membered nitrogen-, oxygen-, and phosphorus-containingheterocyclic ring refers to a 5- to 10-membered nitrogen-, oxygen-, andphosphorus-containing heterocyclic ring which contains only a nitrogenatom, an oxygen atom, and a phosphorus atom as heteroatoms forming thering and which may be fused, such as 1,3,2-oxazaphospholidine,2,3-dihydrobenzo[d][1,3,2]oxazaphosphor, 1,3,2-oxazaphosphinane, or3,4-dihydro-4H-benzo[e][1,3,2]oxazaphosphinine.

The 6- to 10-membered oxygen- and phosphorus-containing heterocyclicring refers to a 6- to 10-membered oxygen- and phosphorus-containingheterocyclic ring which contains only an oxygen atom and a phosphorusatom as heteroatoms forming the ring and which may be fused, such as1,3,2-dioxaphosphinane, 1,3,2-dioxaphosphepane, or1,3,2-dioxaphosphocane.

The C₂₋₆ alkanoyl group refers to a linear or branched C₂₋₆ alkanoylgroup such as an acetyl group, a propionyl group, a valeryl group, anisovaleryl group, or a pivaloyl group.

The C₃₋₈ cycloalkylcarbonyl group refers to a C₃₋₈ cycloalkylcarbonylgroup such as a cyclopropylcarbonyl group, a cyclobutylcarbonyl group, acyclopentylcarbonyl group, a cyclohexylcarbonyl group, or acycloheptylcarbonyl group.

The aroyl group refers to a benzoyl group, a naphthoyl group, or thelike.

The heterocyclic carbonyl group refers to a heterocyclic carbonyl groupsuch as pyrrolylcarbonyl, pyridylcarbonyl, furanylcarbonyl, orthienylcarbonyl.

The acyl group refers to a formyl group, a succinyl group, a glutarylgroup, a maleoyl group, a phthaloyl group, a C₂₋₆ alkanoyl group, a C₃₋₈cycloalkylcarbonyl group, an aroyl group, or a heterocyclic carbonylgroup.

The C₂₋₆ alkanoyloxy group refers to a linear or branched C₂₋₆alkanoyloxy group such as an acetyloxy group, a propionyloxy group, avaleryloxy group, an isovaleryloxy group, or a pivaloyloxy group.

The C₁₋₆ alkylcarbonyloxy group refers to a linear or branched C₁₋₆alkylcarbonyloxy group such as a methylcarbonyloxy group, anethylcarbonyloxy group, a propylcarbonyloxy group, anisopropylcarbonyloxy group, a butylcarbonyloxy group, asec-butylcarbonyloxy group, an isobutylcarbonyloxy group, or atert-butylcarbonyloxy group.

The C₃₋₈ cycloalkylcarbonyloxy group refers to a C₃₋₈cycloalkylcarbonyloxy group such as a cyclopropylcarbonyloxy group, acyclobutylcarbonyloxy group, a cyclopentylcarbonyloxy group, acyclohexylcarbonyloxy group, or a cycloheptylcarbonyloxy group.

The aroyloxy group refers to a benzoyloxy group, a naphthoyloxy group,or the like.

The heterocyclic carbonyloxy group refers to a heterocyclic carbonyloxygroup such as pyrrolylcarbonyloxy, pyridylcarbonyloxy,furanylcarbonyloxy, or thienylcarbonyloxy.

The acyloxy group refers to a C₂₋₆ alkanoyloxy group, a C₃₋₈cycloalkylcarbonyloxy group, an aroyloxy group, or a heterocycliccarbonyloxy group.

The C₂₋₆ alkanoylthio group refers to a linear or branched C₂₋₆alkanoylthio group such as an acetylthio group, a propionylthio group, avalerylthio group, an isovalerylthio group, or a pivaloylthio group.

The C₁₋₆ alkylcarbonylthio group refers to a linear or branched C₁₋₆alkylcarbonylthio group such as a methylcarbonylthio group, anethylcarbonylthio group, a propylcarbonylthio group, anisopropylcarbonylthio group, a butylcarbonylthio group, asec-butylcarbonylthio group, an isobutylcarbonylthio group, or atert-butylcarbonylthio group.

The C₃₋₈ cycloalkylcarbonylthio group refers to a C₃₋₈cycloalkylcarbonylthio group such as a cyclopropylcarbonylthio group, acyclobutylcarbonylthio group, a cyclopentylcarbonylthio group, acyclohexylcarbonylthio group, or a cycloheptylcarbonylthio group.

The aroylthio group refers to a benzoylthio group, a naphthoylthiogroup, or the like.

The heterocyclic carbonylthio group refers to a heterocycliccarbonylthio group such as pyrrolylcarbonylthio, pyridylcarbonylthio,furanylcarbonylthio, or thienylcarbonylthio.

The acylthio group refers to a C₂₋₆ alkanoylthio group, a C₃₋₈cycloalkylcarbonylthio group, an aroylthio group, or a heterocycliccarbonylthio group.

The silyl group refers to trimethylsilyl, triethylsilyl, a tributylsilylgroup, or tert-butylmethylsilyl.

The leaving group refers to a halogen atom, a C₁₋₆ alkylsulfonyloxygroup, an aryloxy group, or an arylsulfonyloxy group. The C₁₋₆alkylsulfonyloxy group, aryloxy group, and arylsulfonyloxy group may besubstituted with one or more substituents selected from a halogen atom,a nitro group, a C₁₋₆ alkyl group, and a C₁₋₆ alkoxy group.

The hydroxyl protecting group is any conventional group which can beused as a protecting group for a hydroxyl group, and examples thereofinclude the groups described in, for example, T. W. Greene et al.,Protective Groups in Organic Synthesis, 4th edition, pp. 16 to 299,2007, John Wiley & Sons, Inc. Specific examples of the hydroxylprotecting group include a C₁₋₆ alkyl group, a C₂₋₆ alkenyl group, anar-C₁₋₆ alkyl group, a C₁₋₆ alkoxy C₁₋₆ alkyl group, an ar-C₁₋₆ alkoxyC₁₋₆ alkyl group, an acyl group, a C₁₋₆ alkoxycarbonyl group, an ar-C₁₋₆alkoxycarbonyl group, a C₁₋₆ alkylsulfonyl group, an arylsulfonyl group,a silyl group, a tetrahydrofuranyl group, and a tetrahydropyranyl group.

The thiol protecting group is any conventional group which can be usedas a protecting group for a thiol group, and examples thereof includethe groups described in, for example, W. Greene et al., ProtectiveGroups in Organic Synthesis, 4th edition, pp. 647 to 695, 2007, JohnWiley & Sons, Inc. Specific examples of the thiol protecting groupinclude a C₁₋₆ alkyl group, a C₂₋₆ alkenyl group, an ar-C₁₋₆ alkylgroup, a C₁₋₆ alkoxy C₁₋₆ alkyl group, an acyl group, and a silyl group.

The amino protecting group is any conventional group which can be usedas a protecting group for an amino group, and examples thereof includethe groups described in, for example, W. Greene et al., ProtectiveGroups in Organic Synthesis, 4th edition, pp. 696 to 926, 2007, JohnWiley & Sons, Inc. Specific examples of the amino protecting groupinclude an ar-C₁₋₆ alkyl group, a C₁₋₆ alkoxy C₁₋₆ alkyl group, an acylgroup, a C₁₋₆ alkoxycarbonyl group, an ar-C₁₋₆ alkoxycarbonyl group, anaryloxycarbonyl group, a C₁₋₆ alkylsulfonyl group, an arylsulfonylgroup, and a silyl group.

Aliphatic hydrocarbons refer to pentane, hexane, heptane, cyclohexane,methylcyclohexane, and ethylcyclohexane.

Halogenated hydrocarbons refer to dichloromethane, chloroform, anddichloroethane.

Ethers refer to diethyl ether, diisopropyl ether, dioxane,tetrahydrofuran, anisole, ethylene glycol dimethyl ether, diethyleneglycol dimethyl ether, and diethylene glycol diethyl ether.

Ketones refer to acetone, 2-butanone, 4-methyl-2-pentanone, and methylisobutyl ketone.

Esters refer to methyl acetate, ethyl acetate, propyl acetate, and butylacetate.

Amides refer to N,N-dimethylformamide, N,N-dimethylacetamide, andN-methylpyrrolidone.

Nitriles refer to acetonitrile and propionitrile.

Sulfoxides refer to dimethyl sulfoxide and sulfolane.

Aromatic hydrocarbons refer to benzene, toluene, and xylene.

The inorganic base refers to sodium hydroxide, potassium hydroxide,sodium methoxide, sodium ethoxide, tert-butoxy sodium, tert-butoxypotassium, sodium hydrogen carbonate, sodium carbonate, potassiumcarbonate, tripotassium phosphate, potassium acetate, cesium fluoride,cesium carbonate, or tert-butyl magnesium chloride.

The organic base refers to triethylamine, N,N-diisopropylethylamine,1,8-diazabicyclo(5.4.0)undec-7-ene (DBU), pyridine,4-dimethylaminopyridine, N-methylmorpholine, or imidazole.

Individual substituent groups have the following meanings.

<Substituent group A>a halogen atom; a hydroxyl group which may beprotected; a cyano group; a nitro group; a carbamoyl group; an oxogroup; a C₁₋₆ alkyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₁₋₆ alkyldisulfanylgroup which may be substituted with one or more substituents selectedfrom Substituent group B; a C₂₋₆ alkenyl group which may be substitutedwith one or more substituents selected from Substituent group B; a C₂₋₆alkynyl group which may be substituted with one or more substituentsselected from Substituent group B; a C₃₋₈ cycloalkyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₃₋₈ cycloalkyldisulfanyl group which may be substituted withone or more substituents selected from Substituent group B; a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group B; a C₁₋₆ alkoxycarbonyloxy group whichmay be substituted with one or more substituents selected fromSubstituent group B; a C₃₋₈ cycloalkoxycarbonyl group which may besubstituted with one or more substituents selected from Substituentgroup B; an aryl group which may be substituted with one or moresubstituents selected from Substituent group B; an aryldisulfanyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a heterocyclic ring group which may be substitutedwith one or more substituents selected from Substituent group B; aheterocyclic oxy group which may be substituted with one or moresubstituents selected from Substituent group B; an acyloxy group whichmay be substituted with one or more substituents selected fromSubstituent group B; an acylthio group which may be substituted with oneor more substituents selected from Substituent group B; anaminocarbonyloxy group which may be substituted with one or moresubstituents selected from Substituent group B; and an aminocarbonylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group B.

<Substituent group B>a halogen atom; a hydroxyl group; a cyano group; anitro group; a carboxyl group; a carbamoyl group; a hydroxymethyl group;a C₁₋₆ alkyl group; a C₂₋₆ alkenyl group; a C₂₋₆ alkynyl group; a C₃₋₈cycloalkyl group; a C₁₋₆ alkoxy group; a C₁₋₆ alkoxycarbonyl group; aC₃₋₈ cycloalkoxycarbonyl group; an ar-C₁₋₆ alkoxycarbonyl group; an arylgroup; an aryloxy group; a heterocyclic oxy group which may besubstituted with one or more substituents selected from a hydroxyl groupand a hydroxymethyl group; an ar-C₁₋₆ alkoxy group; and an acyloxygroup.

Examples of salts of the compound represented by General Formula [1]include salts in basic groups such as an amino group, and salts inacidic groups such as a hydroxyl group and a carboxyl group, which arecommonly known.

Examples of salts in basic groups include salts with mineral acids suchas hydrochloric acid, hydrobromic acid, nitric acid, and sulfuric acid;salts with organic carboxylic acids such as formic acid, acetic acid,citric acid, oxalic acid, fumaric acid, maleic acid, succinic acid,malic acid, tartaric acid, aspartic acid, trichloroacetic acid, andtrifluoroacetic acid; and salts with sulfonic acids such asmethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,mesitylene sulfonic acid, and naphthalene sulfonic acid.

Examples of salts in acidic groups include salts with alkali metals suchas sodium and potassium; salts with alkaline earth metals such ascalcium and magnesium; ammonium salts; and salts withnitrogen-containing organic bases such as trimethylamine, triethylamine,tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine,N-methylmorpholine, diethylamine, dicyclohexylamine, procaine,dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, and N,N′-dibenzylethylenediamine

Among the salts mentioned above, preferred salts includepharmacologically acceptable salts.

The compound represented by General Formula [1] or a salt thereofaccording to the embodiment of the present invention can be used for thetreatment of adenovirus.

The treatment refers to preventing, treating, or the like of a varietyof diseases.

The treatment agent refers to a substance which is provided for thepurpose of preventing or treating a variety of diseases.

The preventing refers to inhibition of disease onset, reduction ofdisease onset risk, delay of disease onset, or the like.

The treating refers to improvement of, inhibition of progression of, orthe like of a target disease or condition.

The compound represented by General Formula [1] according to theembodiment of the present invention is represented by

(in the formula, R¹, R², R³, and R⁴ are as defined above).

R¹

R¹ is a halogen atom, an amino group which may be substituted, a C₁₋₆alkoxy group which may be substituted, a C₃₋₈ cycloalkoxy group whichmay be substituted, a C₁₋₆ alkylthio group which may be substituted, ahydroxyl group which may be protected, or a thiol group which may beprotected.

R¹ is preferably a halogen atom, an amino group which may be substitutedwith one or more substituents selected from Substituent group A, a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, a C₃₋₈ cycloalkoxy group which may besubstituted with one or more substituents selected from Substituentgroup A, a C₁₋₆ alkylthio group which may be substituted with one ormore substituents selected from Substituent group A, a hydroxyl groupwhich may be protected, or a thiol group which may be protected; morepreferably a halogen atom, an amino group which may be substituted withone or more substituents selected from Substituent group A, a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, a hydroxyl group which may beprotected, or a thiol group which may be protected; still morepreferably a halogen atom, an amino group which may be substituted withone or more substituents selected from Substituent group A, a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, or a hydroxyl group; and even stillmore preferably a halogen atom or a C₁₋₆ alkoxy group.

The substituent of the amino group which may be substituted with one ormore substituents selected from Substituent group A in the definition ofR¹ is preferably a C₁₋₆ alkyl group which may be substituted with one ormore substituents selected from Substituent group B or a C₃₋₈ cycloalkylgroup which may be substituted with one or more substituents selectedfrom Substituent group B, and more preferably a C₁₋₆ alkyl group or aC₃₋₈ cycloalkyl group.

The substituent of the C₁₋₆ alkoxy group which may be substituted withone or more substituents selected from Substituent group A in thedefinition of R¹ is preferably a halogen atom.

The substituent of the C₃₋₈ cycloalkoxy group which may be substitutedwith one or more substituents selected from Substituent group A in thedefinition of R¹ is preferably a halogen atom or a C₁₋₆ alkyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B.

The substituent of the C₁₋₆ alkylthio group which may be substitutedwith one or more substituents selected from Substituent group A in thedefinition of R¹ is preferably a halogen atom or a C₁₋₆ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group B.

R²

R² is a hydrogen atom or an amino protecting group.

R² is preferably a hydrogen atom or an acyl group, more preferably ahydrogen atom or a C₂₋₆ alkanoyl group, and still more preferably ahydrogen atom.

R³

R³ is a C₁₋₂₀ alkoxy group which may be substituted, a C₃₋₈ cycloalkoxygroup which may be substituted, a C₁₋₂₀ alkylthio group which may besubstituted, an aryloxy group which may be substituted, a heterocyclicring group which may be substituted, a heterocyclic oxy group which maybe substituted, an amino group which may be substituted, or—O—P(O)(OH)—O—PO₃H, provided that R² is a hydrogen atom in a case whereR³ is —O—P(O)(OH)—O—PO₃H.

R³ is preferably a C₁₋₂₀ alkoxy group which may be substituted with oneor more substituents selected from Substituent group A, a C₃₋₈cycloalkoxy group which may be substituted with one or more substituentsselected from Substituent group A, a C₁₋₂₀ alkylthio group which may besubstituted with one or more substituents selected from Substituentgroup A, an aryloxy group which may be substituted with one or moresubstituents selected from Substituent group A, a heterocyclic ringgroup which may be substituted with one or more substituents selectedfrom Substituent group A, a heterocyclic oxy group which may besubstituted with one or more substituents selected from Substituentgroup A, an amino group which may be substituted with one or moresubstituents selected from Substituent group A, or —O—P(O)(OH)—O—PO₃H;more preferably a C₁₋₂₀ alkoxy group which may be substituted with oneor more substituents selected from Substituent group A, a C₁₋₂₀alkylthio group which may be substituted with one or more substituentsselected from Substituent group A, an aryloxy group which may besubstituted with one or more substituents selected from Substituentgroup A, an amino group which may be substituted with one or moresubstituents selected from Substituent group A, or —O—P(O)(OH)—O—PO₃H;still more preferably a C₁₋₂₀ alkoxy group which may be substituted withone or more substituents selected from Substituent group A, a C₁₋₂₀alkylthio group which may be substituted with one or more substituentsselected from Substituent group A, an aryloxy group which may besubstituted with one or more substituents selected from Substituentgroup A, or —O—P(O)(OH)—O—PO₃H; and even still more preferably a C₁₋₂₀alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, an aryloxy group which may besubstituted with one or more substituents selected from Substituentgroup A, or —O—P(O)(OH)—O—PO₃H.

The substituent of the C₁₋₂₀ alkoxy group which may be substituted withone or more substituents selected from Substituent group A in thedefinition of R³ is preferably a C₁₋₆ alkyldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B, a C₁₋₆ alkylcarbonyloxy group which may be substituted with oneor more substituents selected from Substituent group B, or a C₁₋₆alkylcarbonylthio group which may be substituted with one or moresubstituents selected from Substituent group B.

Here, the substituent of the C₁₋₆ alkyldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B is preferably a hydroxyl group, a C₁₋₆ alkoxy group, an ar-C₁₋₆alkoxy group, or an acyloxy group and more preferably an ar-C₁₋₆ alkoxygroup.

The substituent of the C₁₋₆ alkylcarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B is preferably a hydroxyl group, a C₁₋₆ alkoxy group, an ar-C₁₋₆alkoxy group, or an acyloxy group and more preferably a hydroxyl group.

The substituent of the C₁₋₆ alkylcarbonylthio group which may besubstituted with one or more substituents selected from Substituentgroup B is preferably a hydroxyl group, a C₁₋₆ alkoxy group, an ar-C₁₋₆alkoxy group, or an acyloxy group and more preferably a hydroxyl group.

The substituent of the aryloxy group which may be substituted with oneor more substituents selected from Substituent group A in the definitionof R³ is preferably a halogen atom, a C₁₋₆ alkyl group which may besubstituted with one or more substituents selected from Substituentgroup B, or a C₁₋₆ alkoxy group which may be substituted with one ormore substituents selected from Substituent group B, and more preferablya halogen atom.

R⁴

R⁴ is a C₁₋₂₀ alkoxy group which may be substituted, a C₃₋₈ cycloalkoxygroup which may be substituted, a C₁₋₂₀ alkylthio group which may besubstituted, an aryloxy group which may be substituted, a heterocyclicring group which may be substituted, a heterocyclic oxy group which maybe substituted, an amino group which may be substituted, or a hydroxylgroup which may be protected.

R⁴ is preferably a C₁₋₂₀ alkoxy group which may be substituted with oneor more substituents selected from Substituent group A, a C₃₋₈cycloalkoxy group which may be substituted with one or more substituentsselected from Substituent group A, a C₁₋₂₀ alkylthio group which may besubstituted with one or more substituents selected from Substituentgroup A, an aryloxy group which may be substituted with one or moresubstituents selected from Substituent group A, a heterocyclic ringgroup which may be substituted with one or more substituents selectedfrom Substituent group A, a heterocyclic oxy group which may besubstituted with one or more substituents selected from Substituentgroup A, an amino group which may be substituted with one or moresubstituents selected from Substituent group A, or a hydroxyl groupwhich may be protected; more preferably a C₁₋₂₀ alkoxy group which maybe substituted with one or more substituents selected from Substituentgroup A, a C₁₋₂₀ alkylthio group which may be substituted with one ormore substituents selected from Substituent group A, an aryloxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, an amino group which may be substituted with one ormore substituents selected from Substituent group A, or a hydroxyl groupwhich may be protected; and still more preferably a C₁₋₂₀ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, an amino group which may be substituted with one ormore substituents selected from Substituent group A, or a hydroxyl groupwhich may be protected.

The substituent of the C₁₋₂₀ alkoxy group which may be substituted withone or more substituents selected from Substituent group A in thedefinition of R⁴ is preferably a C₁₋₆ alkyldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B, a C₁₋₆ alkylcarbonyloxy group which may be substituted with oneor more substituents selected from Substituent group B, or a C₁₋₆alkylcarbonylthio group which may be substituted with one or moresubstituents selected from Substituent group B.

Here, the substituent of the C₁₋₆ alkyldisulfanyl group which may besubstituted with one or more substituents selected from Substituentgroup B is preferably a hydroxyl group, a C₁₋₆ alkoxy group, an ar-C₁₋₆alkoxy group, or an acyloxy group and more preferably an ar-C₁₋₆ alkoxygroup.

The substituent of the C₁₋₆ alkylcarbonyloxy group which may besubstituted with one or more substituents selected from Substituentgroup B is preferably a hydroxyl group, a C₁₋₆ alkoxy group, an ar-C₁₋₆alkoxy group, or an acyloxy group and more preferably a hydroxyl group.

The substituent of the C₁₋₆ alkylcarbonylthio group which may besubstituted with one or more substituents selected from Substituentgroup B is preferably a hydroxyl group, a C₁₋₆ alkoxy group, an ar-C₁₋₆alkoxy group, or an acyloxy group and more preferably a hydroxyl group.

The substituent of the aryloxy group which may be substituted with oneor more substituents selected from Substituent group A in the definitionof R⁴ is preferably a halogen atom, a C₁₋₆ alkyl group which may besubstituted with one or more substituents selected from Substituentgroup B, or a C₁₋₆ alkoxy group which may be substituted with one ormore substituents selected from Substituent group B, and more preferablya halogen atom.

R³ and R⁴, together with the phosphorus atom to which R³ and R⁴ arebonded, may be combined to form a 5- to 10-membered nitrogen- andphosphorus-containing heterocyclic ring which may be substituted, a 5-to 10-membered oxygen- and phosphorus-containing heterocyclic ring whichmay be substituted, or a 5- to 10-membered nitrogen-, oxygen-, andphosphorus-containing heterocyclic ring which may be substituted.

The ring formed by combining R³ and R⁴ together with the phosphorus atomto which R³ and R⁴ are bonded is preferably a 5- to 10-membered oxygen-and phosphorus-containing heterocyclic ring which may be substituted andmore preferably 1,3,2-dioxaphosphinane or4H-benzo[d][1,3,2]dioxaphosphinine.

The substituent of the 5- to 10-membered nitrogen- andphosphorus-containing heterocyclic ring which may be substituted, the 5-to 10-membered oxygen- and phosphorus-containing heterocyclic ring whichmay be substituted, or the 5- to 10-membered nitrogen-, oxygen-, andphosphorus-containing heterocyclic ring which may be substituted, eachof which being formed by combining R³ and R⁴ together with thephosphorus atom to which R³ and R⁴ are bonded, is preferably a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group A or an aryl group which may besubstituted with one or more substituents selected from Substituentgroup A.

Here, the substituent of the aryl group which may be substituted withone or more substituents selected from Substituent group A is preferablya halogen atom.

The compound represented by General Formula [1] is preferably a compoundin which R¹ is a halogen atom, an amino group which may be substitutedwith one or more substituents selected from Substituent group A, a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, a hydroxyl group which may beprotected, or a thiol group which may be protected; R² is a hydrogenatom; R³ is a C₁₋₂₀ alkoxy group which may be substituted with one ormore substituents selected from Substituent group A, a C₁₋₂₀ alkylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group A, an aryloxy group which may be substituted withone or more substituents selected from Substituent group A, an aminogroup which may be substituted with one or more substituents selectedfrom Substituent group A, or —O—P(O)(OH)—O—PO₃H; and R⁴ is a C₁₋₂₀alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, a C₁₋₂₀ alkylthio group which may besubstituted with one or more substituents selected from Substituentgroup A, an aryloxy group which may be substituted with one or moresubstituents selected from Substituent group A, an amino group which maybe substituted with one or more substituents selected from Substituentgroup A, or a hydroxyl group which may be protected.

The compound represented by General Formula [1] is more preferably acompound in which R¹ is a halogen atom, an amino group which may besubstituted with one or more substituents selected from Substituentgroup A, a C₁₋₆ alkoxy group which may be substituted with one or moresubstituents selected from Substituent group A, or a hydroxyl group; R²is a hydrogen atom; R³ is a C₁₋₂₀ alkoxy group which may be substitutedwith one or more substituents selected from Substituent group A, a C₁₋₂₀alkylthio group which may be substituted with one or more substituentsselected from Substituent group A, an aryloxy group which may besubstituted with one or more substituents selected from Substituentgroup A, or —O—P(O)(OH)—O—PO₃H; and R⁴ is a C₁₋₂₀ alkoxy group which maybe substituted with one or more substituents selected from Substituentgroup A, an amino group which may be substituted with one or moresubstituents selected from Substituent group A, or a hydroxyl groupwhich may be protected.

The compound represented by General Formula [1] is still more preferablya compound in which R¹ is a halogen atom or a C₁₋₆ alkoxy group; R² is ahydrogen atom; R³ is a C₁₋₂₀ alkoxy group which may be substituted withone or more substituents selected from Substituent group A, an aryloxygroup which may be substituted with one or more substituents selectedfrom Substituent group A, or —O—P(O)(OH)—O—PO₃H; and R⁴ is a C₁₋₂₀alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, an amino group which may besubstituted with one or more substituents selected from Substituentgroup A, or a hydroxyl group which may be protected.

The compound represented by General Formula [1] is preferably methyl

(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)-L-alaninate(Example 3-2-2), methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(naphthalen-1-yloxy)phosphoryl)-L-alaninate(Example 1-2-1), methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chlorophenoxy)phosphoryl)-L-alaninate(Example 1-2-2), methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate(Example 1-1), ethyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate(Example 1-2-7), ethyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chlorophenoxy)phosphoryl)-L-alaninate(Example 1-2-8), methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-bromophenoxy)phosphoryl)-L-alaninate(Example 1-2-9), methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-chlorophenoxy)phosphoryl)-L-alaninate(Example 1-2-10), methyl(((ci-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-(pivaloylthio)ethoxy)phosphoryl)-L-alaninate(Example 1-2-11), methyl(((cis-3-(2-amino-6-ethoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate(Example 1-2-19), methyl2-((((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)amino)-2-methylpropanoate(Example 1-2-25),(cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methylbis(pivaloyloxymethyl)phosphate (Example 5-2), methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chloro-2-fluorophenoxy)phosphoryl)-L-alaninate(Example 1-2-31), or methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(3-bromophenoxy)phosphoryl)-L-alaninate(Example 1-2-32).

In a case where isomers (for example, a tautomer, an optical isomer, anda geometric isomer) are present for the compound represented by GeneralFormula [1] or a salt thereof, the present invention also includes thoseisomers and further includes solvates, hydrates, and various forms ofcrystals.

Next, a method for producing the compound represented by General Formula[1] will be described.

The compound represented by General Formula [1] is produced by combiningmethods known per se, and can be produced, for example, according to thefollowing production methods.

[Production Method 1]

(In the formulae, X represents a leaving group, and R¹, R², R³, and R⁴are as defined above.)

As a compound of General Formula [A1], for example,2-amino-9-(cis-3-(hydroxymethyl)cyclobutyl)-1,9-dihydro-6H-purin-6-oneis known.

As a compound of General Formula [S1], for example,((chlorophosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropanoate)and 2-chloro-4H-benzo[d][1,3,2]dioxaphosphinine 2-oxide are known.

The compound of General Formula [1] can be produced by reacting thecompound of General Formula [A1] with the compound of General Formula[S1] in the presence of a base.

The solvent used in this reaction is not particularly limited as long asit does not affect the reaction and examples thereof include ethers andamides. These solvents may be used as a mixture thereof.

Preferred examples of the solvent include ethers, with tetrahydrofuranbeing more preferred.

The amount of the solvent to be used is not particularly limited, butmay be 1 to 50-fold amount (v/w) with respect to the compound of GeneralFormula [A1].

The amount of the compound of General Formula [S1] to be used may be 1to 20-fold molar amount and preferably 1 to 10-fold molar amount withrespect to the compound of General Formula [A1].

The base used in this reaction may be, for example, tert-butyl magnesiumchloride.

The amount of the base to be used may be 1 to 5-fold molar amount andpreferably 1 to 2-fold molar amount with respect to the compound ofGeneral Formula [A1].

This reaction may be carried out at −78° C. to 100° C., preferably −78°C. to 40° C. for 30 minutes to 48 hours.

The compound of General Formula [A1] and the compound of General Formula[S1] can be derived into other compounds of General Formula [A1] andother compounds of General Formula [S1], for example, by subjecting themto a reaction known per se such as condensation, addition, oxidation,reduction, rearrangement, substitution, halogenation, dehydration, orhydrolysis, or an appropriate combination of these reactions.

In a case where an amino group, a hydroxyl group, or a carboxyl group ispresent in the compounds of General Formula [A1] and intermediatesthereof, the protecting group for such a group can be appropriatelyrearranged to carry out the reaction. In addition, in a case where twoor more protecting groups are present, a reaction known per se can becarried out to make selective deprotection.

Among the compounds used in the above-mentioned production method, acompound that can take the form of a salt can also be used as a salt.Examples of such a salt include the same salts as the salts of thecompound represented by General Formula [1] according to the embodimentof the present invention described above.

In a case where isomers (for example, a tautomer, an optical isomer, anda geometric isomer) are present for the compounds used in theabove-mentioned production method, these isomers can also be used. Inaddition, in a case where solvates, hydrates, and various forms ofcrystals are present, these solvates, hydrates, and various forms ofcrystals can also be used.

The compound represented by General Formula [1] or a salt thereof can beused as an anti-adenoviral agent or as a medicine for treating anadenovirus infection. Examples of the adenovirus infection includerespiratory infection, pharyngoconjunctival fever, epidemickeratoconjunctivitis, hepatitis, gastroenteritis, cystitis, andencephalitis. The anti-adenoviral agent according to the embodiment ofthe present invention and the medicine for treating an adenovirusinfection according to the embodiment of the present invention can beprovided as a pharmaceutical composition.

In a pharmaceutical composition containing the compound represented byGeneral Formula [1] or a salt thereof according to the embodiment of thepresent invention, an additive commonly used in formulation may beappropriately mixed.

Examples of the additive include an excipient, a disintegrating agent, abinding agent, a lubricant, a taste masking agent, a colorant, aflavoring agent, a surfactant, a coating agent, and a plasticizer.

Examples of the excipient include sugar alcohols such as erythritol,mannitol, xylitol, and sorbitol; sugars such as white sugar, powderedsugar, lactose, and glucose; cyclodextrins such as α-cyclodextrin,β-cyclodextrin, γ-cyclodextrin, hydroxypropyl β-cyclodextrin, and sodiumsulfobutylether β-cyclodextrin; celluloses such as crystalline celluloseand microcrystalline cellulose; and starches such as corn starch, potatostarch, and pregelatinized starch.

Examples of the disintegrating agent include carmellose, carmellosecalcium, croscarmellose sodium, sodium carboxymethyl starch,crospovidone, low-substituted hydroxypropyl cellulose, and a partiallypregelatinized starch.

Examples of the binding agent include hydroxypropyl cellulose,carmellose sodium, and methylcellulose.

Examples of the lubricant include stearic acid, magnesium stearate,calcium stearate, talc, hydrated silicon dioxide, light anhydroussilicic acid, and sucrose fatty acid ester.

Examples of the taste masking agent include aspartame, saccharin,stevia, thaumatin, and acesulfame potassium.

Examples of the colorant include titanium dioxide, iron sesquioxide,yellow ferric oxide, black iron oxide, Food Red No. 102, Food Yellow No.4, and Food Yellow No. 5.

Examples of the flavoring agent include an essential oil such as anorange oil, a lemon oil, a peppermint oil, or a pine oil; an essencesuch as an orange essence or a peppermint essence; a flavor such as acherry flavor, a vanilla flavor, or a fruit flavor; a powder fragrancesuch as an apple micron, a banana micron, a peach micron, a strawberrymicron, or an orange micron; vanillin; and ethyl vanillin.

Examples of the surfactant include sodium lauryl sulfate, dioctyl sodiumsulfosuccinate, polysorbate, and polyoxyethylene hydrogenated castoroil.

Examples of the coating agent include hydroxypropyl methyl cellulose, anaminoalkyl methacrylate copolymer E, an aminoalkyl methacrylatecopolymer RS, ethyl cellulose, cellulose acetate phthalate,hydroxypropyl methyl cellulose phthalate, a methacrylic acid copolymerL, a methacrylic acid copolymer LD, and a methacrylic acid copolymer S.

Examples of the plasticizer include triethyl citrate, macrogol,triacetin, and propylene glycol.

These additives may be used alone or in combination of two or morethereof.

Although the formulation amount of the additives is not particularlylimited, the additives may be suitably formulated such that the effectsthereof are sufficiently exhibited depending on the respective purposes.

The pharmaceutical composition to which an appropriate mixture has beenadded can be orally or parenterally administered according to aconventional method in the form of a tablet, a capsule, a powder, asyrup, a granule, a pill, a suspension, an emulsion, a solution, apowdered preparation, a suppository, an eye drop, a nasal drop, an eardrop, a patch, an ointment, an injection, or the like, and is preferablyparenterally administered in the form of eye drop or the like.

The administration method, dosage, and administration frequency of thecompound represented by General Formula [1] or the salt thereofaccording to the embodiment of the present invention can beappropriately selected depending on the age, body weight, and symptomsof the patient. Typically, for an adult, 0.01 to 1,000 mg/kg/day may beadministered orally or parenterally once or in several divided doses. Itis preferred that 0.01 to 1,000 mg/kg/day is administered parenterallyin the form of eye drop or the like once or in several divided doses.

EXAMPLES

Hereinafter, the present invention will be described with reference toReference Examples and Examples, but the present invention is notlimited thereto.

Unless otherwise specified, purification by column chromatography wascarried out using an automated purification apparatus ISOLERA(manufactured by Biotage AB) or a medium-pressure liquid chromatographYFLC-Wprep2XY.N (manufactured by Yamazen Corporation).

Unless otherwise specified, SNAPKP-Sil Cartridge (manufactured byBiotage AB), or HI-FLASH COLUMN W001, W002, W003, W004, or W005(manufactured by Yamazen Corporation) was used as a carrier in silicagel column chromatography; SNAP KP-NH Cartridge (manufactured by BiotageAB) was used as a carrier in basic silica gel column chromatography; andCHROMATOREX Q-PACK Cartridge (manufactured by Fuji Silysia ChemicalLtd.) was used as a carrier in diol silica gel column chromatography.

In preparative thin layer chromatography, PLC glass plate silica gel F₆₀(manufactured by Merck & Co., Inc.) was used.

The mixing ratio in the eluent was a volume ratio. For example,“hexane:ethyl acetate gradient elution=50:50 to 0:100” means that aneluent of 50% hexane/50% ethyl acetate was finally changed to an eluentof 0% hexane/100% ethyl acetate.

In addition, for example, “hexane:ethyl acetate gradient elution=50:50to 0:100, methanol:ethyl acetate gradient elution=0:100 to 20:80” meansthat an eluent of 50% hexane/50% ethyl acetate was changed to an eluentof 0% hexane/100% ethyl acetate, and then the eluent was switched to aneluent of 0% methanol/100% ethyl acetate and finally changed to aneluent of 20% methanol/80% ethyl acetate.

SFC 30 (manufactured by Waters Corporation) was used for supercriticalfluid chromatography.

MS spectra were measured using an ACQUITY SQD LC/MS System (manufacturedby Waters Corporation, ionization method: Electro Spray Ionization (ESI)method), a Model M-8000 (manufactured by Hitachi, Ltd., ionizationmethod: ESI method), or an LCMS-2010EV (manufactured by ShimadzuCorporation, ionization method: method of carrying out ESI andAtmospheric Pressure Chemical Ionization (APCI) at the same time).

As a microwave reactor, Initiator Sixty (manufactured by Biotage AB) wasused.

NMR spectra were measured using Bruker AV300 (manufactured by BrukerCorporation, 300 MHz) and using tetramethylsilane as an internalstandard, and all δ values were shown in ppm.

The retention time (RT) was measured using SQD (manufactured by WatersCorporation), and was shown in minutes (min).

Column: BEHC 18 1.7 μm, 2.1×30 mm (manufactured by Waters Corporation)

Solvent: liquid A: 0.1% formic acid-water

liquid B: 0.1% formic acid-acetonitrile

Gradient cycle: 0.00 min (liquid A/liquid B=95/5), 2.00 min (liquidA/liquid B=5/95), 3.00 min (liquid A/liquid B=5/95), 3.01 min (liquidA/liquid B=100/0), 3.80 min (liquid A/liquid B=100/0)

Flow rate: 0.5 mL/min

Column temperature: room temperature

Detection wavelength: 254 nm

Abbreviations in Examples have the following meanings.

Boc: tert-butoxycarbonyl

Et: ethyl

HATU: 1-(bis(dimethylamino)methylene)-1H-1,2,3-triazolo[4,5,b]pyridinium3-oxide hexafluorophosphate

M: mol/L

Me: methyl

Pr: propyl

RT (min): retention time (min)

*: bonding position

Reference Example 1

First Step

Under a nitrogen atmosphere, a 2.0 M lithium aluminumhydride/tetrahydrofuran solution (25 mL) was added dropwise underice-cooling to a mixture of ethyl cis-3-aminocyclobutane-1-carboxylatehydrochloride (4.3 g) and tetrahydrofuran (43 mL) which was then stirredat room temperature for 1 hour. Water (4 mL), a 15% aqueous sodiumhydroxide solution (4 mL), and water (12 mL) were added to the reactionsolution, and the resulting solid was filtered off and washed withtetrahydrofuran. The solvent was distilled off under reduced pressure togive (cis-3-aminocyclobutyl)methanol (4.4 g) as a colorless oil.

Second Step

A mixture of (cis-3-aminocyclobutyl)methanol (8.5 g) obtained in thefirst step, 2-amino-4,6-dichloro-5-formamidopyrimidine (17.4 g),N,N-diisopropylethylamine (73 mL), ethanol (255 mL), andN-methyl-2-pyrrolidone (8.5 mL) was stirred with heating under refluxfor 6 hours. The solvent was distilled off under reduced pressure, andthe obtained residue was purified by silica gel column chromatography(methanol:ethyl acetate=5:95) to giveN-(2-amino-4-chloro-6-(((cis-3-(hydroxymethyl)cyclobutyl)aminolpyrimidine-5-formamide (8.0 g) as a yellow solid.

MS (ESI m/z): 272, 274 (M+H)

RT (min): 0.44

Third Step

Triethyl orthoformate (24 mL) and a 4 M hydrochloric acid/dioxanesolution (11 mL) were added to a mixture ofN-(2-amino-4-chloro-6-(((cis-3-(hydroxymethyl)cyclobutyl)aminolpyrimidine-5-formamide (7.9 g) and ethanol (40 mL) which was thenstirred at 53° C. for 2 hours. A 4 M hydrochloric acid/dioxane solution(7.2 mL) was added to the reaction solution which was then stirred at53° C. for 9 hours. Isopropyl alcohol (60 mL) was added to the reactionsolution which was then stirred for 2 hours under ice-cooling, and theresulting solid was collected by filtration. A mixture of the obtainedcrude product (5.8 g), ethanol (35 mL), and water (1.7 mL) was stirredat 50° C. for 30 minutes. After cooling to room temperature, the solidwas collected by filtration and washed with ethanol to give(cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanol hydrochloride(3.5 g) as a pale yellow solid.

MS (ESI m/z): 254, 256 (M+H)

RT (min): 0.61

Reference Example 2-1

A 5 M sodium methoxide/methanol solution (0.5 mL) was added to a mixtureof (cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanolhydrochloride (336 mg) and methanol (4.0 mL) which was then stirred at50° C. for 5 hours. The solvent was distilled off under reducedpressure, and the obtained residue was purified by silica gel columnchromatography (hexane:ethyl acetate=50:50 to 0:100, methanol:ethylacetate=0:100 to 20:80) to give(cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methanol (142 mg) asa white solid.

MS (ESI m/z): 250 (M+H)

RT (min): 0.54

Reference Example 2-2

Cyclopropylamine (50 μL) was added to a mixture of(cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanol hydrochloride(20 mg) and ethanol (0.5 mL) which was then irradiated with microwave(microwave reactor, 120° C., 1 hour, 2.45 GHz, 0 to 240 W). The reactionsolution was purified by silica gel column chromatography (hexane:ethylacetate=50:50 to 0:100, methanol:ethyl acetate=0:100 to 20:80) to give(cis-3-(2-amino-6-(cyclopropylamino)-9H-purin-9-yl)cyclobutyllmethanol(21 mg) as a white solid.

MS (ESI m/z): 275 (M+H)

RT (min): 0.55

Reference Example 2-3

A mixture of (cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanol(0.50 g), water (4.0 mL), and formic acid (4.0 mL) was stirred at 70° C.for 2.5 hours. The solvent was distilled off under reduced pressure, anda 7 M ammonia/methanol solution (10 mL) was added to the obtainedresidue which was then allowed to stand for 12 hours. The solvent wasdistilled off under reduced pressure, methanol (2.0 mL) was added to theobtained residue, and the resulting solid was collected by filtration togive2-amino-9-(cis-3-(hydroxymethyl)cyclobutyl)-1,9-dihydro-6H-purin-6-one(0.36 g) as a white solid.

MS (ESI m/z): 236 (M+H)

RT (min): 0.38

Reference Example 2-4

A 50% aqueous dimethylamine solution (0.1 mL) was added to a mixture of(cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanol hydrochloride(20 mg) and ethanol (0.5 mL) which was then stirred at room temperaturefor 1 hour. The reaction solution was purified by silica gel columnchromatography (hexane:ethyl acetate=50:50 to 0:100, methanol:ethylacetate=0:100 to 20:80) to give(cis-3-(2-amino-6-(dimethylamino)-9H-purin-9-yl)cyclobutyllmethanol (18mg) as a white solid.

MS (ESI m/z): 263 (M+H)

RT (min): 0.54

Reference Example 2-5

A 15% aqueous sodium thiomethoxide solution (0.1 mL) was added to amixture of (cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanolhydrochloride (20 mg) and ethanol (0.5 mL) which was then stirred atroom temperature for 1 hour. The reaction solution was purified bysilica gel column chromatography (hexane:ethyl acetate=50:50 to 0:100,methanol:ethyl acetate=0:100 to 20:80) to give(cis-3-(2-amino-6-(methylthio)-9H-purin-9-yl)cyclobutyllmethanol (14 mg)as a white solid.

MS (ESI m/z): 266 (M+H)

RT (min): 0.68

Reference Example 2-6

A 2 M methylamine/tetrahydrofuran solution (0.2 mL) was added to amixture of (cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanolhydrochloride (20 mg) and ethanol (0.5 mL) which was then irradiatedwith microwave (microwave reactor, 120° C., 1 hour, 2.45 GHz, 0 to 240W). The reaction solution was purified by silica gel columnchromatography (hexane:ethyl acetate=50:50 to 0:100, methanol:ethylacetate=0:100 to 20:80) to give(cis-3-(2-amino-6-(methylamino)-9H-purin-9-yl)cyclobutyl)methanol (17mg) as a white solid.

MS (ESI m/z): 249 (M+H)

RT (min): 0.47

Reference Example 2-7

A 20% sodium ethoxide/ethanol solution (0.1 mL) was added to a mixtureof (cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanolhydrochloride (20 mg) and methanol (0.5 mL) which was then stirred atroom temperature for 18 hours. The reaction solution was purified bysilica gel column chromatography (hexane:ethyl acetate=50:50 to 0:100,methanol:ethyl acetate=0:100 to 20:80) to give(cis-3-(2-amino-6-ethoxy-9H-purin-9-yl)cyclobutyl)methanol (15 mg) as awhite solid.

MS (ESI m/z): 264 (M+H)

RT (min): 0.64

Reference Example 2-8

A 15% aqueous sodium hydrosulfide solution (0.1 mL) was added to amixture of (cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanolhydrochloride (20 mg) and ethanol (0.5 mL) which was then stirred atroom temperature for 18 hours. The reaction solution was purified bysilica gel column chromatography (hexane:ethyl acetate=50:50 to 0:100,methanol:ethyl acetate=0:100 to 20:80) to give(cis-3-(2-amino-6-mercapto-9H-purin-9-yl)cyclobutyl)methanol (15 mg) asa white solid.

MS (ESI m/z): 252 (M+H)

RT (min): 0.47

Reference Example 2-9

A mixture of (cis-3-(2-amino-6-chloro-9H-purin-9-yl)cyclobutyl)methanolhydrochloride (20 mg) and a 7 M ammonia/methanol solution (0.5 mL) wasirradiated with microwave (microwave reactor, 130° C., 3 hours, 2.45GHz, 0 to 240 W). The reaction solution was purified by basic silica gelcolumn chromatography (hexane:ethyl acetate=50:50 to 0:100,methanol:ethyl acetate=0:100 to 20:80) to give(cis-3-(2,6-diamino-9H-purin-9-yl)cyclobutyl)methanol (15 mg) as a whitesolid.

MS (ESI m/z): 235 (M+H)

RT (min): 0.40

Reference Example 3-1

Pivaloyl chloride (1.3 mL) was added dropwise at −78° C. to a mixture of2-mercaptoethanol (0.75 mL), triethylamine (2.0 mL), and methylenechloride (25 mL) which was then stirred at room temperature for 2 hours.Water (20 mL) was added to the reaction solution, and the organic layerwas dried over anhydrous sodium sulfate. The solvent was distilled offunder reduced pressure to give S-(2-hydroxyethyl)2,2-dimethylpropanethioate (1.8 g) as a colorless oil.

¹H-NMR (CDCl₃) δ: 3.76 (t, 2H, J=6.1 Hz), 3.06 (t, 2H, J=6.1 Hz), 1.93(br s, 1H), 1.25 (s, 9H).

Reference Example 3-2

The following compound was obtained in the same manner as in ReferenceExample 3-1.

S-(2-hydroxyethyl) 2-methylpropanethioate

¹H-NMR (CDCl₃) δ: 3.77 (t, 2H, J=6.3 Hz), 3.08 (t, 2H, J=6.3 Hz),2.85-2.70 (m, 1H), 1.89 (br s, 1H), 1.21 (d, 6H, J=6.6 Hz).

Reference Example 3-3

Under a nitrogen atmosphere, 60% sodium hydride (0.13 g) was added to amixture of 2,2′ -disulfanediyldiethanol (0.50 g) and tetrahydrofuran(5.0 mL) which was then stirred at room temperature for 10 minutes.Benzyl bromide (0.38 mL) was added to the reaction solution which wasthen stirred for 3 hours, and water was added thereto, followed byextraction with ethyl acetate. The organic layer was washed with anaqueous saturated sodium chloride solution, the solvent was distilledoff under reduced pressure, and then the obtained residue was purifiedby silica gel column chromatography (hexane:ethyl acetate=80:20 to50:50) to give 2-((2-(benzyloxy)ethyl)disulfanyl)ethanol (0.39 g) as acolorless oil.

¹H-NMR (CDCl₃) δ: 7.39-7.27 (m, 5H), 4.56 (s, 2H), 3.92-3.81 (m, 2H),3.75 (t, 2H, J=6.3 Hz), 2.94 (t, 2H, J=6.3 Hz), 2.83 (t, 2H, J=5.9 Hz),2.12-1.95 (m, 1H).

MS (ESI m/z): 245 (M+H)

RT (min): 1.30

Reference Example 4

First Step

Triethylamine (2 mL) and HATU (4.87 g) were added to a mixture ofN-(cert-butoxycarbonyl)-L-alanine (2.5 g), 2-naphthol (1.7 g), andmethylene chloride (12 mL) which was then stirred overnight. Water wasadded to the reaction solution, extraction was carried out with ethylacetate, the solvent was distilled off under reduced pressure, and thenthe obtained residue was purified by silica gel column chromatography(hexane:ethyl acetate=100:0 to 60:40) to give naphthalen-2-yl(tert-butoxycarbonyl)-L-alaninate (0.70 g).

MS (ESI m/z): 316 (M+H)

RT (min): 1.68

Second Step

A mixture of naphthalen-2-yl (cert-butoxycarbonyl)-L-alaninate (0.70 g)and a 4 M hydrochloric acid/cyclopentyl methyl ether solution (5 mL) wasstirred at room temperature overnight. After distilling off the solventunder reduced pressure, ethyl acetate was added to the obtained residueand the resulting solid was collected by filtration to givenaphthalen-2-yl L-alaninate hydrochloride (0.26 g) as a white solid.

MS (ESI m/z): 216 (M+H)

RT (min): 0.81

Reference Example 5-1

Under a nitrogen atmosphere, a mixture of 4-bromophenol (1.2 g),triethylamine (0.94 mL), and methylene chloride (15 mL) was addeddropwise at −78° C. to a mixture of 4-nitrophenyl phosphorodichloridate(1.7 g) and methylene chloride (15 mL) which was then stirred at −78° C.for 30 minutes. A mixture of L-alanine methyl ester hydrochloride (0.94g), triethylamine (1.9 mL), and methylene chloride (30 mL) was addeddropwise at −78° C. to the reaction solution which was then stirred at−78° C. for 30 minutes and at room temperature for 1 hour. Afterdistilling off the solvent under reduced pressure, methylene chloride(10 mL) was added to the obtained residue, and the resulting solid wasfiltered off. After distilling off the solvent under reduced pressure,the obtained residue was purified by silica gel column chromatography(hexane:ethyl acetate=90:10 to 0:100) to give((4-bromophenoxy)(4-nitrophenoxy)phosphoryl)-L-alaninate (1.7 g) as awhite solid.

MS (ESI m/z): 459, 461 (M+H)

RT (min): 1.56

Reference Example 5-2

The compounds in Table 1 were obtained in the same manner as inReference Example 5-1.

TABLE 1

Reference MS Example (ESI m/z) RT No. R Compound name (M + H) (min)5-2-1 

Methyl ((naphthalen-1-yloxy)(4- nitrophenoxy)phosphoryl)-L- alaninate 4

1.5

5-2-2 

Methyl ((4-chlorophenoxy) (4-nitrophenoxy) phosphoryl)-L-alaninate 415417 1.

2 5-2-3 

Methyl (((4-bromonaphthalen-1-yl)oxy) (4-nitrophenoxy)phosphoryl)-L-alaninate

1.75 5-2-4 

Methyl (((4-chloronaphthalen-1-yl) oxy)(4-nitrophenoxy) phosphoryl)-L-alaninate 465 467 1.72 5-2-5 

Ethyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L- alaninate 395 1.495-2-6 

Ethyl ((naphthalen-1-yloxy)(4- nitrophenoxy)phosphoryl)-L- alaninate 4

5 1.65 5-2-7 

Ethyl ((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)- L-alaninate 476 4781.53 5-2-8 

Ethyl ((4-chlorophenoxy) (4-nitrophenoxy) phosphoryl)-L-alaninate 429

1.

1 5-2-9 

Methyl ((2-bromophenoxy)(4- nitrophenoxy)phosphoryl)-L- alaninate

1.

5-2-10

Methyl ((2-chlorophenoxy)(4- nitrophenoxy) phosphoryl)-L-alaninate 4

1.4

5-2-11

Methyl ((4-nitrophenoxy) (2-(pivaloylthio) ethoxy)phosphoryl)-L-alaninate 4

1.

5-2-12

Methyl ((2-isobutyrylthio) ethoxy)(4- nitrophenoxy)phosphoryl)-L-alaninate

1.49 5-2-13

Methyl ((4-bromophenoxy)(4- nitrophenoxy)phosphoryl) glycinate 4

1.48 5-2-14

Methyl ((4-bromophenoxy) (4-nitrophenoxy) phosphoryl)-D-alaninate 4

1.56 5-2-15

Dimethyl ((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)-L- alaninate

1.5

5-2-16

Methyl 2-(((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)amino-2-methylpropanoate 4

1.

5-2-17

Methyl ((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)-L- phenylalaninate

1.

5-2-18

Dipentyl ((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)-L- aspartate

2.14 5-2-19

Methyl ((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)-L- leucinate 5

5

1.77 5-2-20

Methyl N-((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)-N-methyl-L-alaninate 473 475 1.70 5-2-21

4-bromophenyl (4-nitrophenyl)((S)-1-(methyl- amino)-1-oxopropan-2-yl)phosphoramidate 4

1.31 5-2-22

Methyl ((4-chloro-2-fluorophenoxy) (4-nitrophenoxy)phosphoryl)-L-alaninate 432

1.56 5-2-23

Methyl ((3-bromophenoxy) (4-nitrophenoxy) phosphoryl)-L-alaninate 459461 1.

5-2-24

Isobutyl ((4-bromophenoxy) (4-nitrophenoxy) phosphoryl)-L-alaninate 501503 1.

0 5-2-25

Cyclobutyl ((4-bromophenoxy) (4-nitrophenoxy) phosphoryl)-L-alaninate500 502 1.7

5-2-26

Naphthalen-2-yl ((4-bromophenoxy)(4- nitrophenoxy)phosphoryl)-L-alaninate 511 513 1.86

indicates data missing or illegible when filed

Reference Example 6-1

Under a nitrogen atmosphere, triethylamine (0.45 mL) was added to amixture of 4-nitrophenyl phosphorodichloridate (0.16 g) and methylenechloride (2 mL) under ice-cooling, and then2-((2-(benzyloxy)ethyl)disulfanyl)ethanol (0.39 g) and methylenechloride (2 mL) were added thereto, which was followed by stirring atroom temperature for 1.5 hours. The reaction solution was purified bysilica gel column chromatography (hexane:ethyl acetate=80:20 to 25:75)to givebis(2-((2-(benzyloxy)ethyl)disulfanyl)ethyl)(4-nitrophenyl)phosphate(0.33 g) as a colorless oil.

¹H-NMR (CDCl₃) δ: 8.27-8.17 (m, 2H), 7.43-7.24 (m, 10H), 4.54 (s, 4H),4.46-4.34 (m, 4H), 3.79-3.66 (m, 4H), 2.99-2.87 (m, 8H).

Reference Example 6-2

The compounds in Table 2 were obtained in the same manner as inReference Example 6-1.

TABLE 2

Reference MS Example (ESI m/z) RT No. R Compound name (M + H) (min)6-2-1

Bis(S-pivaloyl-2-mercapto- ethan-1-yl)(4-nitrophenyl) phosphate

1.

6-2-2

Bis(S-isobutyroyl-2-mercapto- ethan-1-yl)(4-nitrophenyl) phosphate 4

1.

indicates data missing or illegible when filed

Reference Example 7-1

Under a nitrogen atmosphere, a mixture of perfluorophenol (0.62 g),triethylamine (0.47 mL), and methylene chloride (5 mL) was addeddropwise to a mixture of phenyl phosphorodichloridate (0.5 mL) andmethylene chloride (5 mL) at −78° C., which was followed by stirring at−78° C. for 1 hour. A mixture of S-(2-hydroxyethyl)2,2-dimethylpropanethioate (0.54 g), triethylamine (0.94 mL), andmethylene chloride (5 ml) was added dropwise to the reaction solution at−78° C., which was followed by stirring at −78° C. for 1.5 hours andthen at room temperature for 40 minutes. After distilling off thesolvent under reduced pressure, the obtained residue was purified bysilica gel column chromatography (hexane:ethyl acetate=90:10 to 70:30)to give S-(2-(((perfluorophenoxy)(phenoxy)phosphoryl)oxy)ethyl)2,2-dimethylpropanethioate (0.86 g) as a colorless oil.

MS (ESI m/z): 485 (M+H)

RT (min): 2.01

Reference Example 7-2

The compounds in Table 3 were obtained in the same manner as inReference Example 7-1.

TABLE 3

Reference MS Example (ESI m/z) RT No. R Compound name (M + H) (min)7-2-1

Benzyl ((perfluorophenoxy)(phenoxy) phosphoryl)-L-alaninate 502 1.847-2-2

Isopropyl ((perfluorophenoxy)(phenoxy) phosphoryl)-L-alaninate 454 1.

9 7-2-3

Methyl ((perfluorophenoxy)(phenoxy) phosphoryl)-L-alaninate 426 1.

9 7-2-4

Isopropyl ((naphthalen-1-yloxy) (perfluorophenoxy)phosphory

504 1.

2

indicates data missing or illegible when filed

Reference Example 8

The following compound was obtained according to the method described inChem. Eur. J. 2011, 17, 1649-1659.

2-chloro-4H-benzo[d][1,3,2]dioxaphosphinine 2-oxide

MS (ESI m/z): 205, 207 (M+H)

RT (min): 1.11

Reference Example 9

The following compound was obtained according to the method described inOrg. Lett., Vol. 6, No. 10, 2004, 1555-1556.

((Chlorophosphoryl)bis (oxy))bis (methylene)bis (2,2-dimethylpropanoate)

Example 1-1

Under a nitrogen atmosphere, a 1.0 M tert-butyl magnesiumchloride/tetrahydrofuran solution (0.25 mL) was added dropwise to amixture of (cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methanol(20 mg) and tetrahydrofuran (0.5 mL) which was then stirred at roomtemperature for 30 minutes. A mixture of (methyl((4-bromophenoxy)(4-nitrophenoxy)phosphoryl)-L-alaninate (39 mg) andtetrahydrofuran (0.5 mL) was added to the reaction solution which wasthen stirred at room temperature for 16 hours. An aqueous saturatedammonium chloride solution was added to the reaction solution, which wasfollowed by extraction with ethyl acetate and washing with an aqueoussaturated sodium chloride solution. The solvent was distilled off underreduced pressure, and the obtained residue was purified by silica gelcolumn chromatography (hexane:ethyl acetate=50:50 to 0:100,methanol:ethyl acetate=0:100 to 20:80) to give methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate(23 mg) as a pale yellow solid.

¹H-NMR (MeOD) δ: 7.97-7.86 (m, 1H), 7.36-7.26 (m, 2H), 7.26-7.14 (m,2H), 4.85-4.71 (m, 1H), 4.29-4.12 (m, 2H), 4.09-3.89 (m, 4H), 3.71-3.60(m, 3H), 2.68-2.40 (m, 5H), 1.40-1.28 (m, 3H).

MS (ESI m/z): 569, 571 (M+H).

RT (min): 1.18

Example 1-2

The compounds in Table 4-1 to Table 4-4 were obtained in the same manneras in Example 1-1.

TABLE 4-1

MS (ESI m/z) Example (M + RT No. R X Compound name ¹H-NMR H) (min)1-2-1 

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (naphthalen-1-yloxy) phosphoryl)- L-alanin ¹H-NMR (MeOD) δ:8.20-8.09 (m, 1H), 7.91- 7.72 (m, 2H), 7.72-7.59 (m, 1H), 7.56-7.34 (m,4H), 4.80-4.64 (m, 1H), 4.31-4.16 (m, 2H), 4.16-3.96 (m, 4H), 3.62 (s,3H), 2.64-2.27 (m, 5H), 1.40-1.26 (m, 3H). 541 1.18 1-2-2 

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy)(4-chloro- phenoxy)phosphoryl)- L-alaninate ¹H-NMR (MeOD) δ:7.96-7.88 (m, 1H), 7.37- 7.26 (m, 2H), 7.25-7.14 (m, 2H), 4.85-4.73 (m,1H), 4.28-4.13 (m, 2H), 4.08-3.89 (m, 4H), 3.70-3.60 (m 3H), 2.69-2.39(m, 5H), 1.42-1.24 (m, 3H). 525 527 1.15 1-2-3 

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H-purin- 9-yl)cyclobutyl)methoxy)((4- bromonaphthalen-1-yl) oxy)phosphoryl)- L-alaninate ¹H-NMR(CDCl₃) δ: 8.28-8.10 (m, 2H), 7.75- 7.54 (m, 4H), 7.49-7.41 (m, 1H),4.99 (s, 2H), 4.80-4.66 (m, 1H), 4.36-4.27 (m, 2H), 4.21-4.04 (m, 4H),3.85-3.60 (m, 4H), 2.69-2.47 (m, 5H), 1.46-1.33 (m, 3H). 619 621 1.351-2-4 

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl)methoxy)((4- chloronaphthalen-1-yl) oxy)phosphoryl)-L-alaninate ¹H-NMR (CDCl₃) δ: 8.27-8.22 (m, 1H), 8.17- 8.12 (m, 1H),7.67-7.55 (m, 3H), 7.49 (s, 2H), 4.98 (s, 2H), 4.80-4.65 (m, 1H),4.36-4.25 (m, 2H), 4.20-4.04 (m, 4H), 3.82-3.62 (m, 4H), 2.66-2.47 (m,5H), 1.41-1.33 (m, 3H). 575 577 1.33 1-2-5 

OMe Ethyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (phenoxy)phosphoryl)-L- alaninate ¹H-NMR (CDCl₃) δ: 7.61-7.57(m, 1H), 7.37- 7.12 (m, 5H), 5.03-4.91 (m, 2H), 4.81-4.66 (m, 1H),4.35-3.97 (m, 8H), 3.65-3.52 (m, 1H), 2.68-2.45 (m, 5H), 1.47-1.36 (m,3H), 1.33-1.20 (m, 3H). 505 1.11 1-2-6 

OMe Ethyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy)(naphthalen- 1-yloxy)phosphoryl)- L-alaninate ¹H-NMR (CDCl₃) δ:8.17-8.09 (m, 1H), 7.87- 7.81 (m, 1H), 7.70-7.63 (m, 1H), 7.57-7.48 (m,4H), 7.43-7.35 (m, 1H), 5.01-4.91 (m, 2H), 4.76-4.40 (m, 1H), 4.36-4.22(m, 2H), 4.19-4.00 (m, 6H), 3.78-3.59 (m, 1H), 2.60-2.43 (m, 5H),1.45-1.33 (m, 3H), 1.30-1.14 (m, 3H). 555 1.26 1-2-7 

OMe Ethyl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4- bromophenoxy)phosphoryl)- L-alaninate ¹H-NMR(CDCl₃) δ: 7.67-7.62 (m, 1H), 7.47- 7.38 (m, 2H), 7.19-7.08 (m, 2H),5.04-4.94 (m, 2H), 4.82-4.68 (m, 1H), 4.34-3.93 (m, 8H), 3.76-3.57 (m,1H), 2.71-2.45 (m, 5H), 1.50-1.34 (m, 3H), 1.32-1.20 (m, 3H). 583 5851.26 1-2-8 

OMe Ethyl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4- chlorophenoxy)phosphoryl)- L-alaninate ¹H-NMR(CDCl₃) δ: 7.66-7.62 (m, 1H), 7.31- 7.25 (m, 2H), 7.22-7.14 (m, 2H),5.01-4.93 (m, 2H), 4.81-4.68 (m, 1H), 4.30-4.10 (m, 4H), 4.10-3.95 (m,4H), 3.67-3.54 (m, 1H), 2.67-2.48 (m, 4H), 1.44-1.35 (m, 3H), 1.30-1.21(m, 4H). 539 541 1.24 1-2-9 

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2- bromophenoxy)phosphoryl)- L-alaninate ¹H-NMR(CDCl₃) δ: 7.64-7.49 (m, 3H), 7.32- 7.23 (m, 1H), 7.07-7.00 (m, 1H),5.02-4.91 (m, 2H), 4.80-4.67 (m, 1H), 4.36-4.04 (m, 6H), 3.77-3.64 (m,4H), 2.67-2.42 (m, 5H), 1.47-1.34 (m, 3H). 569 571 1.12 1.13 1-2-10

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2- chlorophenoxy)phosphoryl)- L-alaninate ¹H-NMR(CDCl₃) δ: 7.63-7.59 (m, 1H), 7.55- 7.48 (m, 1H), 7.44-7.37 (m, 1H),7.26-7.18 (m, 1H), 7.14-7.05 (m, 1H), 5.01-4.92 (m, 2H), 4.81-4.65 (m,1H), 4.35-4.03 (m, 6H), 3.79-3.63 (m, 4H), 2.68-2.42 (m, 5H), 1.48-1.33(m, 3H). 525 527 1.10 1.11 1-2-11

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2- (pivaloylthio)ethoxy) phosphoryl)-L-alaninate1H-NMR (CDCl3) δ: 7.71-7.68 (m, 1H), 5.09- 4.95 (m, 2H), 4.83-4.69 (m,1H), 4.25-3.91 (m, 8H), 3.80-3.71 (m, 3H), 3.53-3.37 (m, 1H), 3.24-3.09(m, 2H), 2.72-2.47 (m, 5H), 1.49-1.38 (m, 3H), 1.32-1.20 (m, 9H). 5591.23

TABLE 4-2

MS (ESI m/z) Example (M + RT No. R X Compound name ¹H-NMR H) (min)1-2-12

OMe Methyl (((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (2-(isobutyrylthio) ethoxy)phosphoryl)- L-alaninate¹H-NMR (CDCl₃) δ: 7.72-7.68 (m, 1H), 5.01 (br s, 2H), 4.83-4.69 (m, 1H),4.23-3.91 (m, 8H), 3.78-3.71 (m, 3H), 3.52-3.37 (m, 1H), 3.23- 3.12 (m,2H), 2.83-2.48 (m, 6H), 1.50-1.39 (m, 3H), 1.39-1.22 (m, 6H). 545 1.141-2-13

OMe (cis-3-(2-amino- 6-methoxy-9H- purin-9-yl) cyclobutyl)methylbis(S-isobutyroyl- 2-mercaptoethan- 1-yl)phosphate ¹H-NMR (CDCl₃) δ:7.68-7.64 (m, 1H), 5.07 (br s, 2H), 4.81-4.69 (m, 1H), 4.26-4.17 (m,2H), 4.26-4.10 (m, 4H), 4.06 (s, 3H), 3.18 (t, 4H, J = 6.6 Hz),2.82-2.50 (m, 7H), 1.19 (d, 12H, J = 6.6 Hz). 590 1.46 1-2-14

OMe (cis-3-(2-amino- 6-methoxy-9H- purin-9-yl) cyclobutyl)methylbis(2-((2- (benzyloxy)ethyl) disulfanyl)ethyl) phosphate ¹H-NMR (MeOD)δ: 7.89 (s, 1H), 7.36-7.13 (m, 10H), 4.84-4.69 (m, 1H), 4.54-4.42 (m,4H), 4.35-4.11 (m, 6H), 4.02 (s, 3H), 3.77-3.60 (m, 4H), 3.03-2.79 (m,8H), 2.65-2.45 (m, 5H). 782 1.77 1-2-15

OMe (cis-3-(2-amino- 6-methoxy-9H- purin-9-yl) cyclobutyl)methylbis(S-pivaloyl-2- mercaptoethan-1- yl)phosphate ¹H-NMR (MeOD) δ: 7.95(s, 1H), 4.87-4.78 (m, 1H), 4.27-4.18 (m, 2H), 4.18-4.07 (m, 4H), 4.04(s, 3H), 3.32-3.11 (m, 4H), 2.68-2.55 (m, 5H), 1.21 (s, 18H). 618 1.601-2-16

NMe₂ Methyl (((cis-3-(2-amino- 6-(dimethylamino)- 9H-purin-9-yl)cyclobutyl) methoxy)(4- bromophenoxy) phosphoryl)- L-alaninate ¹H-NMR(MeOD) δ: 7.84-7.78 (m, 1H), 7.52- 7.43 (m, 2H), 7.21-7.11 (m, 2H),4.82-4.69 (m, 1H), 4.26-4.14 (m, 2H), 4.04-3.90 (m, 1H), 3.71-3.62 (m,3H), 3.41 (s, 6H), 2.68-2.31 (m, 5H), 1.41-1.31 (m, 3H). 582 584 1.091-2-17

SMe Methyl (((cis-3-(2- amino-6- (methylthio)-9H- purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl)-L- alaninate ¹H-NMR(MeOD) δ: 8.01-7.95 (m, 1H), 7.51- 7.43 (m, 2H), 7.20-7.10 (m, 2H),4.86-4.77 (m, 1H), 4.29-4.15 (m, 2H), 4.03-3.90 (m, 1H), 3.71-3.63 (m,3H), 2.68-2.43 (m, 8H), 1.39- 1.31 (m, 3H). 585 587 1.28 1-2-18

NHMe Methyl (((cis-3-(2-amino-6- (methylamino)- 9H-purin-9-yl)cyclobutyl) methoxy)(4- bromo- phenoxy) phosphoryl)- L-alaninate ¹H-NMR(MeOD) δ: 7.87-7.80 (m, 1H), 7.51- 7.43 (m, 2H), 7.20-7.11 (m, 2H),4.81-4.70 (m, 1H), 4.29-4.14 (m, 2H), 4.04-3.89 (m, 1H), 3.70-3.65 (m,3H), 3.07- 3.01 (m, 3H), 2.69-2.29 (m, 5H), 1.42-1.31 (m, 3H). 568 5701.04 1-2-19

OEt Methyl (((cis-3-(2-amino- 6-ethoxy-9H- purin-9-yl) cyclobutyl)methoxy)(4- bromophenoxy) phosphoryl)-L- alaninate ¹H-NMR (MeOD) δ:7.96-7.89 (m, 1H), 7.51- 7.43 (m, 2H), 7.19-7.11 (m, 2H), 4.84-4.75 (m,1H), 4.53 (q, 2H, J = 7.0 Hz), 4.27-4.16 (m, 2H), 4.03-3.90 (m, 1H),3.70- 3.65 (m, 3H), 2.69-2.35 (m, 5H), 1.43 (t, 3H, J = 7.0 Hz),1.38-1.32 (m, 3H). 583 585 1.25 1-2-20

NH₂ Methyl ((4-bromophenoxy) ((cis-3-(2,6- diamino-9H-purin-9-yl)cyclobutyl) methoxy) phosphoryl)- L-alaninate ¹H-NMR (MeOD) δ:7.92-7.85 (m, 1H), 7.51- 7.44 (m, 2H), 7.21-7.11 (m, 2H), 4.83-4.71 (m,2H), 4.27-4.15 (m, 2H), 4.03-3.91 (m, 1H), 3.71-3.65 (m, 3H), 2.69- 2.38(m, 5H), 1.39-1.31 (m, 3H). 554 556 1.02 1-2-21

Cl Methyl (((cis-3-(2-amino- 6-chloro-9H- purin-9-yl) cyclobutyl)methoxy)(4- bromophenoxy) phosphoryl)-L- alaninate ¹H-NMR (MeOD) δ:8.17-8.13 (m, 1H), 7.51- 7.44 (m, 2H), 7.20-7.11 (m, 2H), 4.91-4.84 (m,1H), 4.28-4.16 (m, 2H), 4.03-3.90 (m, 1H), 3.69-3.66 (m, 3H), 2.67- 2.54(m, 5H), 1.39-1.31 (m, 3H). 573 575 1.25 1-2-22

OMe Methyl (((cis-3-(2-amino- 6-methoxy- 9H-purin-9-yl) cyclobutyl)methoxy)(4- bromophenoxy) phosphoryl) glycinate ¹H-NMR (MeOD) δ: 7.66(s, 1H), 7.47-7.40 (m, 2H), 7.18-7.10 (m, 2H), 4.99 (s, 2H), 4.82-4.69(m, 1H), 4.33-4.24 (m, 2H), 4.07 (s, 3H), 3.86- 3.73 (m, 5H), 3.68-3.57(m, 1H), 2.68-2.48 (m, 5H). 555 557 1.11

TABLE 4-3

MS Example (ESI m/z) RT No. R X Compound name ¹H-NMR (M + H) (min)1-2-23

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy)(4- bromophenoxy) phosphoryl)-D- alaninate ¹H-NMR (CDCl₃) δ:7.68-7.64 (m, 1H), 7.49- 7.39 (m, 2H), 7.18-7.08 (m, 2H), 5.01 (s, 2H),4.84-4.69 (m, 1H), 4.35-3.98 (m, 6H), 3.78-3.62 (m, 4H), 2.72-2.45 (m,5H), 1.47-1.35 (m, 3H). 569 571 1.17 1-2-24

OMe Dimethyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl)-L- aspartate ¹H-NMR (CDCl₃) δ:7.69-7.63 (m, 1H), 7.47- 7.39 (m, 2H), 7.17-7.08 (m, 2H), 5.01 (s, 2H),4.82-4.67 (m, 1H), 4.36-4.19 (m, 3H), 4.12-3.98 (m, 4H), 3.74-3.71 (m,3H), 3.66-3.63 (m, 3H), 3.01-2.44 (m, 7H). 627 629 1.17 1-2-25

OMe Methyl 2-((((cis-3-(2-amino- 6-methoxy- 9H-purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl) amino)-2-methylpropanoate ¹H-NMR (CDCl₃) δ: 7.64 (s, 1H), 7.47-7.38 (m, 2H),7.18-7.10 (m, 2H), 4.98 (s, 2H), 4.83-4.68 (m, 1H), 4.28-4.21 (m, 2H),4.07 (s, 3H), 4.01- 3.94 (m, 1H), 3.73 (s, 3H), 2.68-2.47 (m, 5H), 1.56(s, 3H), 1.54 (s, 3H). 583 585 1.23 1-2-26

OMe Methyl (((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl)-L- phenylalaninate ¹H-NMR (CDCl₃)δ: 7.63 (s, 1H), 7.44-7.34 (m, 2H), 7.29-7.18 (m, 3H), 7.13-6.99 (m,4H), 4.96 (s, 2H), 4.79-4.65 (m, 1H), 4.32-4.19 (m, 1H), 4.17-3.81 (m,5H), 3.71-3.65 (m, 3H), 3.47-3.33 (m, 1H), 3.10-2.92 (m, 2H), 2.64-2.34(m, 5H). 645 647 1.40 1-2-27

OMe Dipentyl (((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl)-L- aspartate ¹H-NMR (CDCl₃) δ:7.68-7.61 (m, 1H), 7.46- 7.39 (m, 2H), 7.17-7.08 (m, 2H), 4.99 (s, 2H),4.82-4.67 (m, 1H), 4.34-3.95 (m, 11H), 3.00- 2.46 (m, 7H), 1.71-1.50 (m,4H), 1.39-1.19 (m, 8H), 0.93-0.84 (m, 6H). 739 741 1.84 1-2-28

OMe Methyl (((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy)(4- bromophenoxy) phosphoryl)-L- leucinate ¹H-NMR (CDCl₃) δ:7.68-7.62 (m, 1H), 7.47- 7.39 (m, 2H), 7.17-7.08 (m, 2H), 4.99 (s, 2H),4.82-4.67 (m, 1H), 4.31-4.18 (m, 2H), 4.07 (s, 3H), 4.05-3.88 (m, 1H),3.71-3.65 (m, 3H), 3.47-3.30 (m, 1H), 2.67-2.45 (m, 5H), 1.76-1.41 (m,3H), 0.93-0.84 (m, 6H). 611 613 1.42 1-2-29

OMe Methyl N-(((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl)-N- methyl-L-alaninate ¹H-NMR(CDCl₃) δ: 7.77-7.66 (m, 1H), 7.47- 7.39 (m, 2H), 7.18-7.19 (m, 2H),4.99-4.91 (m, 2H), 4.87-4.71 (m, 1H), 4.62-4.37 (m, 1H), 4.33-4.13 (m,2H), 4.07 (s, 3H), 3.72-3.62 (m, 3H), 2.78-2.45 (m, 8H), 1.42-1.27 (m,3H). 583 585 1.28 1.30 1-2-30

OMe (cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl) methyl)(4-bromophenyl)((S)- 1-(methylamino)- 1-oxopropan-2-yl) phosphoramidate¹H-NMR (CDCl₃) δ: 7.64-7.60 (m, 1H), 7.48- 7.39 (m, 2H), 7.16-7.09 (m,2H), 6.19-6.10 (m, 1H), 5.07-4.96 (m, 2H), 4.80-4.68 (m, 1H), 4.29-4.23(m, 2H), 4.07 (s, 3H), 3.92-3.78 (m, 1H), 3.64-3.54 (m, 1H), 2.77-2.72(m, 3H), 2.68-2.48 (m, 5H), 1.40-1.34 (m, 3H). 568 570 0.98 1-2-31

OMe Methyl (((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy)(4-chloro- 2-fluorophenoxy) phosphoryl)- L-alaninate ¹H-NMR(CDCl₃) δ: 7.68-7.65 (m, 1H), 7.43- 7.34 (m, 1H), 7.19-7.12 (m, 1H),7.11-7.05 (m, 1H), 4.97 (s, 2H), 4.83-4.69 (m, 1H), 4.34-4.23 (m, 2H),4.17-4.00 (m, 4H), 3.76-3.61 (m, 4H), 2.69-2.47 (m, 5H), 1.45-1.36 (m,3H). 543 545 1.18 1-2-32

OMe Methyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy)(3- bromophenoxy) phosphoryl)-L- alaninate ¹H-NMR (CDCl₃) δ:7.64-7.60 (m, 1H), 7.44- 7.39 (m, 1H), 7.32-7.26 (m, 1H), 7.23-7.14 (m,2H), 4.98 (s, 2H), 4.82-4.68 (m, 1H), 4.34-4.16 (m, 2H), 4.15-3.96 (m,4H), 3.75-3.71 (m, 3H), 3.70-3.55 (m, 1H), 2.68-2.48 (m, 5H), 1.44-1.37(m, 3H). 569 571 1.17 1-2-33

OMe Isobutyl (((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy)(4- bromophenoxy) phosphoryl)-L- alaninate ¹H-NMR (CDCl₃) δ:7.65-7.62 (m, 1H), 7.46- 7.38 (m, 2H), 7.16-7.08 (m, 2H), 4.98 (br s,2H), 4.80-4.67 (m, 1H), 4.31-4.20 (m, 2H), 4.10-3.96 (m, 4H), 3.95-3.81(m, 2H), 3.69-3.55 (m, 1H), 2.67-2.49 (m, 5H), 1.98-1.83 (m, 1H),1.45-1.36 (m, 3H), 0.90 (d, 6H, J = 6.6 Hz). 611 613 1.44

TABLE 4-4

MS Example (ESI m/z) RT No. R X Compound name ¹H-NMR (M + H) (min)1-2-34

OMe Cyclobutyl (((cis-3-(2-amino- 6-methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl)-L- alaninate ¹H-NMR (CDCl₃) δ:7.65 (s, 1H), 7.45-7.3

 (m, 2H), 7.16-7.08 (m, 2H), 5.03-4.92 (m, 3H), 4.

1-4.68 (m, 1H), 4.30-4.19 (m, 2H), 4.07 (s, 3H), 4.04-3.92 (m, 1H),3.68-3.54 (m, 1H), 2.67-2.48 (m, 5H), 2.39-2.26 (m, 2H), 2.09-1.95 (m,2H), 1.86-1.66 (m, 2H), 1.42-1.

5 (m, 3H). 509 511 1.38 1-2-35

OMe Naphthalen-2-yl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl) methoxy)(4- bromophenoxy) phosphoryl)-L-alaninate¹H-NMR (CDCl₃) δ: 7.88-7.72 (m, 3H), 7.65- 7.59 (m, 1H), 7.54-7.41 (m,5H), 7.24-7.

 (m, 3H), 5.02-4.90 (m, 2H), 4.

0-4.67 (m, 1H), 4.40-4.26 (m, 3H), 4.08-4.03 (m, 3H), 3.82-3.55 (m, 1H),2.70-2.54 (m, 5H), 1.74-1.59 (m, 3H). 581 583 1.5

indicates data missing or illegible when filed

Example 2-1

Under a nitrogen atmosphere, a 1.0 M tert-butyl magnesiumchloride/tetrahydrofuran solution (0.20 mL) was added dropwise to amixture of2-amino-9-(cis-3-(hydroxymethyl)cyclobutyl)-1,9-dihydro-6H-purin-6-one(20 mg), N,N-dimethylformamide (0.5 mL), and tetrahydrofuran (2.0 mL)which was then stiffed at room temperature for 1 hour. A mixture ofmethyl ((naphthalen-1-yloxy)(4-nitrophenoxy)phosphoryl)-L-alaninate (36mg) and tetrahydrofuran (0.5 mL) was added to the reaction solutionwhich was then stirred at room temperature for 18 hours. An aqueoussaturated ammonium chloride solution was added to the reaction solution,which was followed by extraction with ethyl acetate and washing with anaqueous saturated sodium chloride solution. The solvent was distilledoff under reduced pressure, and the obtained residue was purified bysilica gel column chromatography (hexane:ethyl acetate=50:50 to 0:100,methanol:ethyl acetate=0:100 to 20:80) to give methyl(((cis-3-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)cyclobutyl)methoxy)(naphthalen-1-yloxy)phosphoryl)-L-alaninate (8.5 mg) as a pale yellow solid.

¹H-NMR (MeOD) δ: 8.20-8.11 (m, 1H), 7.90-7.80 (m, 1H), 7.75-7.59 (m,2H), 7.59-7.33 (m, 2H), 4.77-4.62 (m, 1H), 4.29-4.14 (m, 2H), 4.14-3.97(m, 1H), 3.67-3.56 (m, 3H), 2.59-2.28 (m, 5H), 1.40-1.26 (m, 3H).

MS (ESI m/z): 527 (M+H)

RT (min): 1.05

Example 2-2

The compounds in Table 5 were obtained in the same manner as in Example2-1.

TABLE 5

MS Example (ESI m/z) RT No. R X Compound name ¹H-NMR (M + H) (min) 2-2-1

OH Methyl (((cis-3-(2-amino-6- hydroxy-9H- purin-9-yl)cyclobutyl)methoxy)(4- chlorophenoxy) phosphoryl)-L- alaninate ¹H-NMR (MeOD) δ:7.84-7.76 (m, 1H), 7.37- 7.27 (m, 2H), 7.27-7.14 (m, 2H), 4.81-4.69 (m,1H), 4.27-4.14 (m, 2H), 4.03-3.89 (m, 1H), 3.70-3.62 (m, 3H), 2.67-2.37(m, 5H), 1.43-1.25 (m, 3H). 511 513 1.02 2-2-2

OH Methyl (((cis-3-(2-amino- 6-hydroxy-9H- purin-9-yl)cyclobutyl)methoxy)(4- bromophenoxy) phosphoryl)-L- alaninate ¹H-NMR (MeOD) δ:7.85-7.75 (m, 1H), 7.51- 7.42 (m, 2H), 7.20-7.07 (m, 2H), 4.82-4.67 (m,1H), 4.29-4.11 (m, 2H), 4.02-3.86 (m, 1H), 3.72-3.63 (m, 3H), 2.66-2.37(m, 5H), 1.40-1.29 (m, 3H). 555 557 1.04 2-2-3

SH Methyl (((cis-3-(2-amino-6- mercapto-9H-purin-9-yl)cyclobutyl)methoxy) (4-bromophenoxy) phosphoryl)-L- alaninate¹H-NMR (MeOD) δ: 7.96-7.92 (m, 1H), 7.51- 7.43 (m, 2H), 7.19-7.11 (m,2H), 4.82-4.70 (m, 1H), 4.28-4.14 (m, 2H), 4.03-3.89 (m, 1H), 3.70-3.64(m, 3H), 2.67-2.40 (m, 5H), 1.38-1.31 (m, 3H). 571 573 1.11

Example 3-1

Under a nitrogen atmosphere, a 1.0 M tert-butyl magnesiumchloride/tetrahydrofuran solution (0.25 mL) was added dropwise to amixture of (cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methanol(20 mg) and tetrahydrofuran (0.2 mL) which was then stirred at roomtemperature for 20 minutes. A mixture ofS-(2-(((perfluorophenoxy)(phenoxy)phosphoryl)oxy)ethyl)2,2-dimethylpropanethioate (30 mg) and tetrahydrofuran (0.15 mL) wasadded to the reaction solution which was then stirred at roomtemperature for 11 hours. An aqueous saturated ammonium chloridesolution was added to the reaction solution, which was followed byextraction with ethyl acetate and washing with an aqueous saturatedsodium chloride solution. The solvent was distilled off under reducedpressure, and the obtained residue was purified by silica gel columnchromatography (hexane:ethyl acetate=50:50 to 0:100, methanol:ethylacetate=0:100 to 5:95) to giveS-(2-((((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)oxy)ethyl)2,2-dimethylpropanethioate (7.0 mg) as a pale yellow solid.

¹H-NMR (MeOD) δ: 7.89-7.84 (m, 1H), 7.42-7.30 (m, 2H), 7.27-7.17 (m,3H), 4.86-4.74 (m, 1H), 4.40-4.15 (m, 4H), 4.04 (s, 3H), 3.22-3.11 (m,2H), 2.65-2.49 (m, 5H), 1.19 (s, 9H).

MS (ESI m/z): 550 (M+H)

RT (min): 1.45

Example 3-2

The compounds in Table 6 were obtained in the same manner as in Example3-1.

TABLE 6

MS Example (ESI m/z) RT No. R X Compound name ¹H-NMR (M + H) (min) 3-2-1

OMe Benzyl (((cis-3-(2-amino-6- methoxy-9H-purin-9-yl)cyclobutyl)methoxy) (phenoxy)phosphoryl)- L-alaninate ¹H NMR (MeOD)δ: 7.99-7.80 (m, 1H), 7.36- 7.08 (m, 10H), 5.13-5.06 (m, 2H), 4.82-4.65(m, 1H), 4.18-3.94 (m, 6H), 2.59-2.30 (m, 5H), 1.40-1.27 (m, 3H). 5671.30 3-2-2

OMe Methyl (((cis-3-(2-amino- 6-methoxy- 9H-purin-9-yl) cyclobutyl)methoxy)(phenoxy) phosphoryl)- L-alaninate 1H NMR (MeOD) δ: 7.92-7.84(m, 1H), 7.38- 7.09 (m, 5H), 4.90-4.72 (m, 1H), 4.26-4.07 (m, 2H),4.07-3.89 (m, 4H), 3.69-3.60 (m, 3H), 2.66-2.37 (m, 5H), 1.41-1.25 (m,3H). 491 1.03 3-2-3

NHcycloPr Isopropyl (((cis-3-(2-amino-6- (cyclopropyl-amino)-9H-purin-9- yl)cyclobutyl) methoxy)(phenoxy) phosphoryl)-L-alaninate ¹H NMR (MeOD) δ: 7.78 (s, 1H), 7.37-7.11 (m, 5H), 5.00-4.92(m, 1H), 4.81-4.67 (m, 1H), 4.20-4.14 (m, 2H), 3.97-3.83 (m, 1H),2.95-2.85 (m, 1H), 2.64-2.49 (m, 3H), 2.49-2.33 (m, 2H), 1.38-1.31 (m,3H), 1.24-1.17 (m, 6H), 0.88-0.80 (m, 2H), 0.64-0.56 (m, 2H). 544 1.093-2-4

OMe Isopropyl (((cis-3-(2-amino-6- methoxy-9H- purin-9-yl)cyclobutyl)methoxy) (phenoxy)phosphoryl)- L-alaninate ¹H NMR (MeOD) δ: 7.88 (s,1H), 7.38-7.11 (m, 5H), 5.10-4.92 (m, 1H), 4.84-4.72 (m, 1H), 4.22-4.15(m, 2H), 4.04 (s, 3H), 3.97-3.84 (m, 1H), 2.64-2.40 (m, 5H), 1.38-1.31(m, 3H), 1.25-1.17 (m, 6H). 519 1.08 3-2-5

OH Benzyl (((cis-3-(2-amino-6- hydroxy-9H- purin-9-yl)cyclobutyl)methoxy) (phenoxy)phosphoryl)- L-alaninate ¹H NMR (MeOD) δ: 7.77-7.72(m, 1H), 7.38- 7.10 (m, 10H), 5.17-5.08 (m, 2H), 4.79-4.63 (m, 1H),4.18-3.93 (m, 3H), 2.58-2.28 (m, 5H), 1.39-1.30 (m, 3H). 553 1.19 3-2-6

OH Isopropyl (((cis-3-(2-amino-6- hydroxy-9H- purin-9-yl)cyclobutyl)methoxy) (naphthalen-1-yloxy) phosphoryl)- L-alaninate ¹H NMR (MeOD) δ:8.20-8.13 (m, 1H), 7.89- 7.82 (m, 1H), 7.72-7.61 (m, 2H), 7.55-7.36 (m,4H), 5.00-4.83 (m, 1H), 4.74-4.61 (m, 1H), 4.28-4.17 (m, 2H), 4.05-3.93(m, 1H), 2.57-2.27 (m 5H), 1.38-1.30 (m, 3H), 1.22-1.14 (m, 6H). 5551.19 1.18 3-2-7

OH Methyl (((cis-3-(2-amino-6- hydroxy-9H- purin-9-yl)cyclobutyl)methoxy) (phenoxy)phosphoryl)- L-alaninate ¹H NMR (MeOD) δ: 7.76-7.69(m, 1H), 7.38- 7.13 (m, 5H), 4.81-4.67 (m, 1H), 4.20-4.12 (m, 2H),4.03-3.93 (m, 1H), 3.67 (s, 3H), 2.63-2.33 (m, 5H), 1.39-1.33 (m, 3H).477 0.90 3-2-8

OH Isopropyl (((cis-3-(2-amino-6- hydroxy-9H- purin-9-yl)cyclobutyl)methoxy)(phenoxy) phosphoryl)-L- alaninate ¹H NMR (MeOD) δ: 7.75 (s,1H), 7.38-7.12 (m, 5H), 5.01-4.85 (m, 1H), 4.81-4.68 (m, 1H), 4.21-4.13(m, 2H), 3.96-3.84 (m, 1H), 2.61-2.37 (m, 5H), 1.38-1.31 (m, 3H),1.25-1.18 (m, 6H). 505 1.06

Example 4-1

Acetyl chloride (28 μL) was added to a mixture of isopropyl(((cis-3-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)-L-alaninate(7 mg) and pyridine (0.1 mL) which was then stirred at room temperaturefor 5 hours. Water was added to the reaction solution, followed byextraction with ethyl acetate, the solvent was distilled off underreduced pressure, and the obtained residue was purified by silica gelcolumn chromatography (methanol:ethyl acetate=0:100 to 20:80) to giveisopropyl(((cis-3-(2-acetamido-6-oxo-1,6-dihydro-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)-L-alaninate(4.4 mg) as a pale yellow solid.

¹H-NMR (MeOD) δ: 7.96 (s, 1H), 7.38-7.12 (m, 5H), 5.01-4.78 (m, 2H),4.23-4.17 (m, 2H), 3.97-3.84 (m, 1H), 2.63-2.51 (m, 5H), 2.22 (s, 3H),1.38-1.31 (m, 3H), 1.25-1.18 (m, 6H).

MS (ESI m/z): 547 (M+H)

RT (min): 1.20

Example 4-2

The following compound was obtained in the same manner as in Example4-1.

Benzyl

(((cis-3-(2-acetamido-6-oxo-1,6-dihydro-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)-L-alaninate

¹H-NMR (MeOD) δ: 7.97-7.92 (m, 1H), 7.39-7.11 (m, 10H), 5.16-5.07 (m,2H), 4.86-4.75 (m, 1H), 4.24-3.94 (m, 3H), 2.62-2.38 (m, 5H), 2.24-2.18(m, 3H), 1.40-1.31 (m, 3H).

MS (ESI m/z): 595 (M+H)

RT (min): 1.30

Example 5-1

Under a nitrogen atmosphere,(cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methanol (20 mg) andN-methylimidazole (38 μL) were added to a mixture of2-chloro-4H-benzo[d][1,3,2]dioxaphosphinine 2-oxide (49 mg) andacetonitrile (1.0 mL) which was then stirred at room temperature for 1hour. The reaction solution was purified by silica gel columnchromatography (hexane:ethyl acetate=70:30 to 0:100, methanol:ethylacetate=0:100 to 15:85) to give2-((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)-4H-benzo[d][1,3,2]dioxaphosphinine2-oxide (16 mg) as a pale yellow solid.

¹H-NMR (CDCl₃) δ: 7.56 (s, 1H), 7.36-7.29 (m, 1H), 7.18-7.05 (m, 3H),5.48-5.31 (m, 2H), 4.99 (s, 2H), 4.84-4.66 (m, 1H), 4.43-4.31 (m, 2H),4.06 (s, 3H), 2.67-2.51 (m, 5H).

MS (ESI m/z): 418 (M+H)

RT (min): 0.97

Example 5-2

The following compound was obtained in the same manner as in Example5-1.

(cis-3-(2-Amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methylbis(pivaloyloxymethyl)phosphate

¹H-NMR (CDCl₃) δ: 7.64 (s, 1H), 5.72-5.63 (m, 4H), 5.04 (s, 2H),4.78-4.68 (m, 1H), 4.28-4.22 (m, 2H), 4.07 (s, 3H), 2.75-2.50 (m, 5H),1.23 (s, 18H).

MS (ESI m/z): 558 (M+H)

RT (min): 1.42

Example 6

Under a nitrogen atmosphere, a mixture of 3-(hexadecyloxy)propan-1-ol(36 mg) and tetrahydrofuran (0.2 mL) was slowly added dropwise to amixture of phosphorus oxychloride (11 μL), triethylamine (34 μL), andtetrahydrofuran (0.5 mL) under ice-cooling, which was followed bystirring at room temperature for 30 minutes.(cis-3-(2-Amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methanol (10 mg) wasadded to the reaction solution which was then stirred at roomtemperature for 12 hours. Water (0.10 mL) and a 2 M aqueous sodiumhydroxide solution (50 μL) were added to the reaction solution which wasthen stirred at room temperature for 15 minutes. The reaction mixturewas purified by diol silica gel column chromatography (hexane:ethylacetate=80:20 to 0:100, methanol:ethyl acetate=0:100 to 40:60) to give(cis-3-(2-amino-6)-methoxy-9H-purin-9-yl)cyclobutyl)methyl(3-(hexadecyloxy)propyl)hydrogenphosphate (6.3 mg) as a pale yellow solid.

¹H-NMR (CDCl₃) δ: 7.77 (s, 1H), 4.79-4.64 (m, 1H), 4.08-3.90 (m, 5H),3.54-3.41 (m, 2H), 3.39-3.30 (m, 2H), 3.11-3.00 (m, 2H), 2.74-2.39 (m,5H), 1.96-1.82 (m, 2H), 1.57-1.41 (m, 2H), 1.35-1.14 (m, 26H), 0.88 (t,3H, J=6.6 Hz).

MS (ESI m/z): 612 (M+H)

RT (min): 2.37

Example 7

The following compound was obtained according to the method described inNucleosides Nucleotides Nucleic Acids. 2006; 25(4-6): 539-51.

(cis-3-(2-Amino-6-oxo-1,6-dihydro-9H-purin-9-yl)cyclobutyl)methyltetrahydrogen triphosphate

Examples 8-1-1 and 8-1-2

Methyl

(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)-L-alaninateobtained in Example 3-2-2 was subjected to chiral resolution bysupercritical fluid chromatography to give optically active substance Aand optically active substance B.

Example 8-1-1 (Optically Active Substance A)

¹H-NMR (CDCl₃) δ: 7.59 (s, 1H), 7.38-7.12 (m, 5H), 4.96 (br s, 2H),4.82-4.67 (m, 1H), 4.34-4.18 (m, 2H), 4.18-4.04 (m, 4H), 3.71 (s, 3H),3.61-3.48 (m, 1H), 2.70-2.48 (m, 5H), 1.39 (d, 3H, J=7.3 Hz).

MS (ESI m/z): 491 (M+H)

RT (min): 1.01

Example 8-1-2 (Optically Active Substance B)

¹H-NMR (CDCl₃) δ: 7.59 (s, 1H), 7.38-7.12 (m, 5H), 4.95 (br s, 2H),4.82-4.66 (m, 1H), 4.33-4.14 (m, 2H), 4.14-3.97 (m, 4H), 3.72 (s, 3H),3.65-3.52 (m, 1H), 2.68-2.45 (m, 5H), 1.40 (d, 3H, J=6.6 Hz).

MS (ESI m/z): 491 (M+H)

RT (min): 1.02

(Conditions for Supercritical Fluid Chromatography)

Column: CHIRALPAK IC (manufactured by Daicel Chemical Industries, Ltd.)

Mobile phase: carbon dioxide/methanol (volume ratio: 70/30)

Flow rate: 30 mL/min

Detection wavelength: 280 nm

Temperature: 40° C.

Retention time: 8.67 min (optically active substance A), 13.49 min(optically active substance B)

Examples 8-2-1 and 8-2-2

Methyl

(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chlorophenoxy)phosphoryl)-L-alaninateobtained in Example 1-2-2 was subjected to chiral resolution bysupercritical fluid chromatography to give optically active substance Aand optically active substance B.

Example 8-2-1 (Optically Active Substance A)

¹H-NMR (CDCl₃) δ: 7.65 (s, 1H), 7.32-7.24 (m, 2H), 7.21-7.14 (m, 2H),4.97 (br s, 2H), 4.83-4.68 (m, 1H), 4.31-4.22 (m, 2H), 4.13-4.02 (m,4H), 3.71 (s, 3H), 3.62-3.50 (m, 1H), 2.67-2.52 (m, 5H), 1.39 (d, 3H,J=7.3 Hz).

MS (ESI m/z): 525, 527 (M+H)

RT (min): 1.14

Example 8-2-2 (Optically Active Substance B)

¹H-NMR (CDCl₃) δ: 7.64 (s, 1H), 7.34-7.24 (m, 2H), 7.23-7.15 (m, 2H),4.96 (s, 2H), 4.82-4.67 (m, 1H), 4.31-4.18 (m, 2H), 4.12-3.98 (m, 4H),3.72 (s, 3H), 3.66-3.55 (m, 1H), 2.67-2.49 (m, 5H), 1.40 (d, 3H, J=7.3Hz).

MS (ESI m/z): 525, 527 (M+H)

RT (min): 1.15

(Conditions for Supercritical Fluid Chromatography)

Column: CHIRALPAK IC (manufactured by Daicel Chemical Industries, Ltd.)

Mobile phase: carbon dioxide/methanol (volume ratio: 70/30)

Flow rate: 30 mL/min

Detection wavelength: 280 nm

Temperature: 40° C.

Retention time: 7.99 min (optically active substance A), 13.02 min(optically active substance B)

Comparative Example 1 (Reference Example 2-3)

2-Amino-9-(cis-3-(hydroxymethyl)cyclobutyl)-1,9-dihydro-6H-purin-6-one

Test Example 1: Evaluation of Anti-Adenovirus Activity

A system in which CRL-11516 cells (available from ATCC) were infectedwith adenovirus type 6 (hereinafter, referred to as ADV6) wasconstructed, and a test for evaluating the anti-adenovirus activity ofthe compound according to the embodiment of the present invention andthe compound of Comparative Example was carried out using the system.The test for evaluating the anti-adenovirus activity was carried outaccording to the method described below.

CRL-11516 cells were added to a 96-well plate at a cell density of 1×10⁴cells/well and cultured at 37° C. for 24 hours. After the culture wascompleted, the cell culture liquid was removed, and a serial dilution(100 μL) of the compound according to the embodiment of the presentinvention or the compound of Comparative Example was added. ADV6(corresponding to 50TCID₅₀) was added thereto, followed by culturing at37° C. for 48 hours.

After the culture was completed, the infected cells were stained usingAdeno-X Rapid Titer Kit (manufactured by Takara Bio Inc.). 100% methanolwas added thereto and the cells were fixed at −20° C. for 10 minutes.The plate was washed three times with phosphate buffered saline(hereinafter, referred to as PBS) and then Mouse Anti-Hexon Antibody wasadded thereto, followed by incubation at 37° C. for 1 hour. After theincubation was completed, the plate was washed three times with PBS, andRat Anti-Mouse Antibody was added thereto, followed by incubation at 37°C. for 1 hour. After the incubation was completed, a mixed stainingsolution of Stable Peroxidase Buffer:DAB Substrate=10:1 was addedthereto, followed by incubation at room temperature for 10 minutes forstaining of the cells.

The number of dark brown-stained cells was counted under a microscope,and the concentration of the test drug that reduces the darkbrown-stained cells by 50% was defined as EC₅₀. EC₅₀ values werecalculated for the triplicate test results, and a mean value and astandard deviation thereof were determined. The results are shown inTable 7 below.

Evaluation Standards

+++: 0.1 μM>EC₅₀

++: 1 μM>EC₅₀≥0.1 μM

+: 10 μM>EC₅₀≥1 μM

−: IC₅₀≥10 μM

TABLE 7 Anti-AdV activity EC₅₀ (μmol/L) Example No. 1-1 +++ 1-2-1 ++1-2-2 +++ 1-2-15 + 3-1 + 3-2-1 ++ 3-2-2 ++ 3-2-4 + 8-2-1 +++ 8-2-2 ++Comparative Example No. 1 − Evaluation standards +++: 0.1 μM > EC₅₀ ++:1 μM > EC₅₀ ≥ 0.1 μM +: 10 μM > EC₅₀ ≥ 1 μM −: IC₅₀ ≥ 10 μM

The compound according to the embodiment of the present invention had anexcellent anti-adenovirus activity.

Test Example 2: Evaluation of ADV DNA Polymerase Inhibitory Activity

Recombinant ADV DNA polymerase was constructed, and a test forevaluating the DNA polymerase inhibitory activity of the compounddescribed in Example 7 was carried out. The test for evaluating the DNApolymerase inhibitory activity was carried out according to the methoddescribed below.

1) Purification of recombinant ADV DNA polymerase

Full-length human adenovirus 19 DNA polymerase (AFA46720.1) was totallysynthesized as a cDNA optimized for insect cell expression, a His tagsequence was added to the C-terminus thereof, and then the resultingconstruct was inserted into pFastBac™ 1 (available from InvitrogenCorporation).

A bacmid was constructed by introducing the above vector into MAXEfficiency (registered trademark) DH10Bac™ competent cells (availablefrom Invitrogen Corporation). It was confirmed by sequencing that theconstructed bacmid contained the full-length human adenovirus 19 DNApolymerase.

Sf9 cells were transfected with the bacmid using Cellfectin II(available from Invitrogen Corporation). The culture supernatantcontaining recombinant baculoviruses was recovered after 7 days, and anoperation of infecting Sf9 cells with baculoviruses was repeated twiceto obtain a sufficient amount of baculoviruses for protein expression.The baculovirus titer was measured using BacPAK™ Baculovirus Rapid TiterKit (manufactured by Takara Bio Inc.).

0.8 ×10⁶ cells/mL of Sf9 cells were infected with baculoviruses inGrace's Insect medium (manufactured by Gibco Corporation) supplementedwith 10% fetal bovine serum (hereinafter, referred to as FBS,manufactured by Gibco Corporation) and 0.1% F-68 (manufactured by GibcoCorporation), such that the multiplicity of infection was about 0.3.After 3 days, Sf9 cells were recovered and the cell pellet was frozen at−80° C.

The frozen cell pellet was suspended in a cell lysis solution (50 mMsodium phosphate buffer (pH 7.0), 10% glycerol, 300 mM sodium chloride,1% Triton (registered trademark) X-100, cOmplete™ EDTA-free ProteaseInhibitor Cocktail (manufactured by F. Hoffmann-La Roche AG)) which wasthen allowed to stand on ice for 15 minutes. After vortexing for 30seconds, centrifugation was carried out at 15,000 rpm to obtain asupernatant containing human adenovirus 19 DNA polymerase.

The supernatant containing human adenovirus 19 DNA polymerase was addedto TALON (registered trademark) Superflow Metal Affinity Resin(manufactured by Takara Bio Inc.) which was then stirred at 4° C. for2.5 hours with a rotator. The TALON (registered trademark) SuperflowMetal Affinity Resin was washed five times with a wash solution 1 (10 mMimidazole, 50 mM sodium phosphate buffer (pH 7.0), 10% glycerol, 300 mMsodium chloride) and three times with a wash solution 2 (25 mMimidazole, 50 mM sodium phosphate buffer (pH 7.0), 10% glycerol, 300 mMsodium chloride). Then, an eluate (50 mM and 100 mM imidazole, 50 mMsodium phosphate buffer (pH 7.0), 10% glycerol, 300 mM sodium chloride)was added to the TALON (registered trademark) Superflow Metal AffinityResin to elute the human adenovirus 19 DNA polymerase. The eluate wasconcentrated with Amicon Ultra-15 Centrifugal Filter Units, MWCO 50K(manufactured by MilliporeSigma Corporation), and then replaced withstock buffer (50 mM sodium phosphate buffer (pH 7.0), 10% glycerol, 300mM sodium chloride) by a dialysis method using Slide-A-Lyzer MINIDialysis Devices, 20K MWCO (manufactured by Thermo Fisher ScientificInc.) prior to use for testing. The concentration of human adenovirus 19DNA polymerase was measured using Pierce™ BCA Protein Assay Kit.

2) Test for evaluating ADV DNA polymerase inhibitory activity

1.5 μM primer oligo DNA (IRDye800-5′-GTAAAACGACGGCCAGT-3′) (SEQ IDNO: 1) and 1 μM template oligo DNA(5′-CCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTTGGCACTGGCCGTCGTTTTACAACGTCGTGA-3′) (SEQ ID NO: 2) were incubated in annealing buffer(10 mM Tris-HCl buffer (pH 8.0), 1 mM ethylenediaminetetraacetic acid(hereinafter, referred to as EDTA), 50 mM sodium chloride) at 95° C. for5 minutes, and then the temperature was gradually returned to roomtemperature for annealing.

Human adenovirus 19 DNA polymerase, primer oligo DNA/template oligo DNA,dNTPs (manufactured by Nippon Gene Co., Ltd.), and the compounddescribed in Example 7 were diluted in assay buffer (50 mM Tris-HClbuffer (pH 7.5), 1 mM dithiothreitol, 4% glycerol, 5 mM magnesiumchloride, 0.1% bovine serum albumin)

40 nM human adenovirus 19 DNA polymerase (5 μL), the compound (5 μL) ata concentration four times the final concentration and described inExample 7, and 40 μM dNTPs (5 μL) were mixed and incubated at 37° C. for5 minutes.

120 nM/80 nM primer oligo DNA/template oligo DNA (5 μL) was added tostart an enzymatic reaction. After the enzymatic reaction at 37° C. for10 minutes, 20 μL of 2×sample buffer (98% formamide, 10 mM EDTA, 0.2%bromophenol blue) was added to stop the reaction.

After incubating at 70° C. for 10 minutes, a part of the reactionsolution was applied to Novex TBE-Urea gels, 15%, 15 well (manufacturedby Thermo Fisher Scientific Inc.), and DNA was separated byelectrophoresis. The extension of the primer oligo DNA by the DNApolymerase activity was evaluated by detecting IRDye800 with Odyssey(manufactured by LI-COR, Inc.). The results are shown in FIG. 1.

The compound according to the embodiment of the present invention had anexcellent inhibitory activity on the DNA extension reaction by ADV DNApolymerase.

The compound represented by General Formula [1] or a salt thereofaccording to an aspect of the present invention is useful as ananti-adenoviral agent. The compound represented by General Formula [1]or a salt thereof according to the aspect of the present invention isuseful as an agent for treating adenovirus.

SEQUENCE LISTING

International Application 18F01188W1JP19029448_44. app under the PatentCooperation Treaty.

What is claimed is:
 1. A compound represented by General Formula [1]:

in the formula, R¹ represents a halogen atom, an amino group which maybe substituted, a C₁₋₆ alkoxy group which may be substituted, a C₃₋₈cycloalkoxy group which may be substituted, a C₁₋₆ alkylthio group whichmay be substituted, a hydroxyl group which may be protected, or a thiolgroup which may be protected; R² represents a hydrogen atom or an aminoprotecting group; R³ represents a C₁₋₂₀ alkoxy group which may besubstituted, a C₃₋₈ cycloalkoxy group which may be substituted, a C₁₋₂₀alkylthio group which may be substituted, an aryloxy group which may besubstituted, a heterocyclic ring group which may be substituted, aheterocyclic oxy group which may be substituted, an amino group whichmay be substituted, or —O—P(O)(OH)—O—PO₃H; and R⁴ represents a C₁₋₂₀alkoxy group which may be substituted, a C₃₋₈ cycloalkoxy group whichmay be substituted, a C₁₋₂₀ alkylthio group which may be substituted, anaryloxy group which may be substituted, a heterocyclic ring group whichmay be substituted, a heterocyclic oxy group which may be substituted,an amino group which may be substituted, or a hydroxyl group which maybe protected; or R³ and R⁴, together with a phosphorus atom to which R³and R⁴ are bonded, may be combined to form a 5- to 10-membered nitrogen-and phosphorus-containing heterocyclic ring which may be substituted, a5- to 10-membered oxygen- and phosphorus-containing heterocyclic ringwhich may be substituted, or a 5- to 10-membered nitrogen-, oxygen-, andphosphorus-containing heterocyclic ring which may be substituted,provided that R² represents a hydrogen atom in a case where R³represents —O—P(O)(OH)—O—PO₃H; or a salt thereof.
 2. The compoundaccording to claim 1, wherein R² is a hydrogen atom; or the saltthereof.
 3. The compound according to claim 1, wherein R′ is a halogenatom, an amino group which may be substituted with one or moresubstituents selected from Substituent group A, a C₁₋₆ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, a C₃₋₈ cycloalkoxy group which may be substitutedwith one or more substituents selected from Substituent group A, a C₁₋₆alkylthio group which may be substituted with one or more substituentsselected from Substituent group A, a hydroxyl group which may beprotected, or a thiol group which may be protected; or the salt thereof,Substituent group A: a halogen atom; a hydroxyl group which may beprotected; a cyano group; a nitro group; a carbamoyl group; an oxogroup; a C₁₋₆ alkyl group which may be substituted with one or moresubstituents selected from Substituent group B; a C₁₋₆ alkyldisulfanylgroup which may be substituted with one or more substituents selectedfrom Substituent group B; a C₂₋₆ alkenyl group which may be substitutedwith one or more substituents selected from Substituent group B; a C₂₋₆alkynyl group which may be substituted with one or more substituentsselected from Substituent group B; a C₃₋₈ cycloalkyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₃₋₈ cycloalkyldisulfanyl group which may be substituted withone or more substituents selected from Substituent group B; a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group B; a C₁₋₆ alkoxycarbonyloxy group whichmay be substituted with one or more substituents selected fromSubstituent group B; a C₃₋₈ cycloalkoxycarbonyl group which may besubstituted with one or more substituents selected from Substituentgroup B; an aryl group which may be substituted with one or moresubstituents selected from Substituent group B; an aryldisulfanyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a heterocyclic ring group which may be substitutedwith one or more substituents selected from Substituent group B; aheterocyclic oxy group which may be substituted with one or moresubstituents selected from Substituent group B; an acyloxy group whichmay be substituted with one or more substituents selected fromSubstituent group B; an acylthio group which may be substituted with oneor more substituents selected from Substituent group B; anaminocarbonyloxy group which may be substituted with one or moresubstituents selected from Substituent group B; and an aminocarbonylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group B, Substituent group B: a halogen atom; ahydroxyl group; a cyano group; a nitro group; a carboxyl group; acarbamoyl group; a hydroxymethyl group; a C₁₋₆ alkyl group; a C₂₋₆alkenyl group; a C₂₋₆ alkynyl group; a C₃₋₈ cycloalkyl group; a C₁₋₆alkoxy group; a C₁₋₆ alkoxycarbonyl group; a C₃₋₈ cycloalkoxycarbonylgroup; an ar-C₁₋₆ alkoxycarbonyl group; an aryl group; an aryloxy group;a heterocyclic oxy group which may be substituted with one or moresubstituents selected from a hydroxyl group and a hydroxymethyl group;an ar-C₁₋₆ alkoxy group; and an acyloxy group.
 4. The compound accordingto claim 3, wherein R¹ is a halogen atom, an amino group which may besubstituted with one or more substituents selected from Substituentgroup A, a C₁₋₆ alkoxy group which may be substituted with one or moresubstituents selected from Substituent group A, a hydroxyl group whichmay be protected, or a thiol group which may be protected; or the saltthereof.
 5. The compound according to claim 3, wherein R¹ is a halogenatom, an amino group which may be substituted with one or moresubstituents selected from Substituent group A, a C₁₋₆ alkoxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, or a hydroxyl group; or the salt thereof.
 6. Thecompound according to claim 1, wherein R³ is a C₁₋₂₀ alkoxy group whichmay be substituted with one or more substituents selected fromSubstituent group A, a C₃₋₈ cycloalkoxy group which may be substitutedwith one or more substituents selected from Substituent group A, a C₁₋₂₀alkylthio group which may be substituted with one or more substituentsselected from Substituent group A, an aryloxy group which may besubstituted with one or more substituents selected from Substituentgroup A, a heterocyclic ring group which may be substituted with one ormore substituents selected from Substituent group A, a heterocyclic oxygroup which may be substituted with one or more substituents selectedfrom Substituent group A, an amino group which may be substituted withone or more substituents selected from Substituent group A, or—O—P(O)(OH)—O—PO₃H; or the salt thereof, Substituent group A: a halogenatom; a hydroxyl group which may be protected; a cyano group; a nitrogroup; a carbamoyl group; an oxo group; a C₁₋₆ alkyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₁₋₆ alkyldisulfanyl group which may be substituted with oneor more substituents selected from Substituent group B; a C₂₋₆ alkenylgroup which may be substituted with one or more substituents selectedfrom Substituent group B; a C₂₋₆ alkynyl group which may be substitutedwith one or more substituents selected from Substituent group B; a C₃₋₈cycloalkyl group which may be substituted with one or more substituentsselected from Substituent group B; a C₃₋₈ cycloalkyldisulfanyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a C₁₋₆ alkoxy group which may be substituted withone or more substituents selected from Substituent group B; a C₁₋₆alkoxycarbonyloxy group which may be substituted with one or moresubstituents selected from Substituent group B; a C₃₋₈cycloalkoxycarbonyl group which may be substituted with one or moresubstituents selected from Substituent group B; an aryl group which maybe substituted with one or more substituents selected from Substituentgroup B; an aryldisulfanyl group which may be substituted with one ormore substituents selected from Substituent group B; a heterocyclic ringgroup which may be substituted with one or more substituents selectedfrom Substituent group B; a heterocyclic oxy group which may besubstituted with one or more substituents selected from Substituentgroup B; an acyloxy group which may be substituted with one or moresubstituents selected from Substituent group B; an acylthio group whichmay be substituted with one or more substituents selected fromSubstituent group B; an aminocarbonyloxy group which may be substitutedwith one or more substituents selected from Substituent group B; and anaminocarbonylthio group which may be substituted with one or moresubstituents selected from Substituent group B, Substituent group B: ahalogen atom; a hydroxyl group; a cyano group; a nitro group; a carboxylgroup; a carbamoyl group; a hydroxymethyl group; a C₁₋₆ alkyl group; aC₂₋₆ alkenyl group; a C₂₋₆ alkynyl group; a C₃₋₈ cycloalkyl group; aC₁₋₆ alkoxy group; a C₁₋₆ alkoxycarbonyl group; a C₃₋₈cycloalkoxycarbonyl group; an ar-C₁₋₆ alkoxycarbonyl group; an arylgroup; an aryloxy group; a heterocyclic oxy group which may besubstituted with one or more substituents selected from a hydroxyl groupand a hydroxymethyl group; an ar-C₁₋₆ alkoxy group; and an acyloxygroup.
 7. The compound according to claim 6, wherein R³ is a C₁₋₂₀alkoxy group which may be substituted with one or more substituentsselected from Substituent group A, a C₁₋₂₀ alkylthio group which may besubstituted with one or more substituents selected from Substituentgroup A, an aryloxy group which may be substituted with one or moresubstituents selected from Substituent group A, an amino group which maybe substituted with one or more substituents selected from Substituentgroup A, or —O—P(O)(OH)—O—PO₃H; or the salt thereof.
 8. The compoundaccording to claim 6, wherein R³ is a C₁₋₂₀ alkoxy group which may besubstituted with one or more substituents selected from Substituentgroup A, a C₁₋₂₀ alkylthio group which may be substituted with one ormore substituents selected from Substituent group A, an aryloxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, or —O—P(O)(OH)—O—PO₃H; or the salt thereof.
 9. Thecompound according to claim 1, wherein R⁴ is a C₁₋₂₀ alkoxy group whichmay be substituted with one or more substituents selected fromSubstituent group A, a C₃₋₈ cycloalkoxy group which may be substitutedwith one or more substituents selected from Substituent group A, a C₁₋₂₀alkylthio group which may be substituted with one or more substituentsselected from Substituent group A, an aryloxy group which may besubstituted with one or more substituents selected from Substituentgroup A, a heterocyclic ring group which may be substituted with one ormore substituents selected from Substituent group A, a heterocyclic oxygroup which may be substituted with one or more substituents selectedfrom Substituent group A, an amino group which may be substituted withone or more substituents selected from Substituent group A, or ahydroxyl group which may be protected; or the salt thereof, Substituentgroup A: a halogen atom; a hydroxyl group which may be protected; acyano group; a nitro group; a carbamoyl group; an oxo group; a C₁₋₆alkyl group which may be substituted with one or more substituentsselected from Substituent group B; a C₁₋₆ alkyldisulfanyl group whichmay be substituted with one or more substituents selected fromSubstituent group B; a C₂₋₆ alkenyl group which may be substituted withone or more substituents selected from Substituent group B; a C₂₋₆alkynyl group which may be substituted with one or more substituentsselected from Substituent group B; a C₃₋₈ cycloalkyl group which may besubstituted with one or more substituents selected from Substituentgroup B; a C₃₋₈ cycloalkyldisulfanyl group which may be substituted withone or more substituents selected from Substituent group B; a C₁₋₆alkoxy group which may be substituted with one or more substituentsselected from Substituent group B; a C₁₋₆ alkoxycarbonyloxy group whichmay be substituted with one or more substituents selected fromSubstituent group B; a C₃₋₈ cycloalkoxycarbonyl group which may besubstituted with one or more substituents selected from Substituentgroup B; an aryl group which may be substituted with one or moresubstituents selected from Substituent group B; an aryldisulfanyl groupwhich may be substituted with one or more substituents selected fromSubstituent group B; a heterocyclic ring group which may be substitutedwith one or more substituents selected from Substituent group B; aheterocyclic oxy group which may be substituted with one or moresubstituents selected from Substituent group B; an acyloxy group whichmay be substituted with one or more substituents selected fromSubstituent group B; an acylthio group which may be substituted with oneor more substituents selected from Substituent group B; anaminocarbonyloxy group which may be substituted with one or moresubstituents selected from Substituent group B; and an aminocarbonylthiogroup which may be substituted with one or more substituents selectedfrom Substituent group B, Substituent group B: a halogen atom; ahydroxyl group; a cyano group; a nitro group; a carboxyl group; acarbamoyl group; a hydroxymethyl group; a C₁₋₆ alkyl group; a C₂₋₆alkenyl group; a C₂₋₆ alkynyl group; a C₃₋₈ cycloalkyl group; a C₁₋₆alkoxy group; a C₁₋₆ alkoxycarbonyl group; a C₃₋₈ cycloalkoxycarbonylgroup; an ar-C₁₋₆ alkoxycarbonyl group; an aryl group; an aryloxy group;a heterocyclic oxy group which may be substituted with one or moresubstituents selected from a hydroxyl group and a hydroxymethyl group;an ar-C₁₋₆ alkoxy group; and an acyloxy group.
 10. The compoundaccording to claim 9, wherein R⁴ is a C₁₋₂₀ alkoxy group which may besubstituted with one or more substituents selected from Substituentgroup A, a C₁₋₂₀ alkylthio group which may be substituted with one ormore substituents selected from Substituent group A, an aryloxy groupwhich may be substituted with one or more substituents selected fromSubstituent group A, an amino group which may be substituted with one ormore substituents selected from Substituent group A, or a hydroxyl groupwhich may be protected; or the salt thereof.
 11. The compound accordingto claim 9, wherein R⁴ is a C₁₋₂₀ alkoxy group which may be substitutedwith one or more substituents selected from Substituent group A, anamino group which may be substituted with one or more substituentsselected from Substituent group A, or a hydroxyl group which may beprotected; or the salt thereof.
 12. The compound according to claim 1,wherein a ring formed by combining R³ and R⁴ together with a phosphorusatom to which R³ and R⁴ are bonded is a 5- to 10-membered oxygen- andphosphorus-containing heterocyclic ring which may be substituted; or thesalt thereof.
 13. A compound selected from methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(phenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(naphthalen-1-yloxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chlorophenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate,ethyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate,ethyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chlorophenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-bromophenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-chlorophenoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(2-(pivaloylthio)ethoxy)phosphoryl)-L-alaninate,methyl(((cis-3-(2-amino-6-ethoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)-L-alaninate,methyl2-((((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-bromophenoxy)phosphoryl)amino)-2-methylpropanoate,(cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methylbis(pivaloyloxymethyl)phosphate, methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(4-chloro-2-fluorophenoxy)phosphoryl)-L-alaninate,and methyl(((cis-3-(2-amino-6-methoxy-9H-purin-9-yl)cyclobutyl)methoxy)(3-bromophenoxy)phosphoryl)-L-alaninate;or a salt thereof.
 14. An anti-adenoviral agent comprising: the compoundor salt thereof according to claim
 1. 15. A method for suppressingadenovirus, comprising: administering the compound or salt thereofaccording to claim 1.